IL296057A - Ribonucleic acid-guided genome recombination engineering at the scale of thousands of bases - Google Patents
Ribonucleic acid-guided genome recombination engineering at the scale of thousands of basesInfo
- Publication number
- IL296057A IL296057A IL296057A IL29605722A IL296057A IL 296057 A IL296057 A IL 296057A IL 296057 A IL296057 A IL 296057A IL 29605722 A IL29605722 A IL 29605722A IL 296057 A IL296057 A IL 296057A
- Authority
- IL
- Israel
- Prior art keywords
- protein
- sequence
- seq
- aptamer
- nucleic acid
- Prior art date
Links
- 108090000623 proteins and genes Proteins 0.000 claims description 221
- 102000004169 proteins and genes Human genes 0.000 claims description 171
- 210000004027 cell Anatomy 0.000 claims description 113
- 150000007523 nucleic acids Chemical class 0.000 claims description 98
- 108091033409 CRISPR Proteins 0.000 claims description 91
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 77
- 230000006798 recombination Effects 0.000 claims description 63
- 238000005215 recombination Methods 0.000 claims description 63
- 102000039446 nucleic acids Human genes 0.000 claims description 61
- 108020004707 nucleic acids Proteins 0.000 claims description 61
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 59
- 238000000034 method Methods 0.000 claims description 57
- 108091023037 Aptamer Proteins 0.000 claims description 51
- 230000000813 microbial effect Effects 0.000 claims description 47
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 46
- 239000013598 vector Substances 0.000 claims description 41
- 108020005004 Guide RNA Proteins 0.000 claims description 36
- 108091008324 binding proteins Proteins 0.000 claims description 34
- 108020001507 fusion proteins Proteins 0.000 claims description 32
- 102000037865 fusion proteins Human genes 0.000 claims description 32
- 108091008103 RNA aptamers Proteins 0.000 claims description 31
- 239000000203 mixture Substances 0.000 claims description 29
- 102000040430 polynucleotide Human genes 0.000 claims description 23
- 108091033319 polynucleotide Proteins 0.000 claims description 23
- 239000002157 polynucleotide Substances 0.000 claims description 23
- 230000000295 complement effect Effects 0.000 claims description 22
- 108060002716 Exonuclease Proteins 0.000 claims description 21
- 241000282414 Homo sapiens Species 0.000 claims description 21
- 102000013165 exonuclease Human genes 0.000 claims description 21
- 230000007115 recruitment Effects 0.000 claims description 21
- 108010079855 Peptide Aptamers Proteins 0.000 claims description 18
- 101710125418 Major capsid protein Proteins 0.000 claims description 15
- 241000193996 Streptococcus pyogenes Species 0.000 claims description 13
- 150000001413 amino acids Chemical class 0.000 claims description 12
- 210000003527 eukaryotic cell Anatomy 0.000 claims description 12
- 210000005260 human cell Anatomy 0.000 claims description 12
- 230000030648 nucleus localization Effects 0.000 claims description 12
- 101710132601 Capsid protein Proteins 0.000 claims description 11
- 108010008532 Deoxyribonuclease I Proteins 0.000 claims description 11
- 102000007260 Deoxyribonuclease I Human genes 0.000 claims description 11
- 101710141454 Nucleoprotein Proteins 0.000 claims description 11
- 101710094648 Coat protein Proteins 0.000 claims description 9
- 102220605874 Cytosolic arginine sensor for mTORC1 subunit 2_D10A_mutation Human genes 0.000 claims description 9
- 102100021181 Golgi phosphoprotein 3 Human genes 0.000 claims description 9
- 101710083689 Probable capsid protein Proteins 0.000 claims description 9
- 101100107610 Arabidopsis thaliana ABCF4 gene Proteins 0.000 claims description 8
- 102100026662 Delta and Notch-like epidermal growth factor-related receptor Human genes 0.000 claims description 8
- 101000911710 Escherichia phage T7 Exonuclease Proteins 0.000 claims description 8
- 101100068078 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) GCN4 gene Proteins 0.000 claims description 8
- 210000004962 mammalian cell Anatomy 0.000 claims description 8
- 238000001727 in vivo Methods 0.000 claims description 7
- 210000000130 stem cell Anatomy 0.000 claims description 6
- 101100166144 Staphylococcus aureus cas9 gene Proteins 0.000 claims description 5
- 230000004075 alteration Effects 0.000 claims description 2
- 238000002054 transplantation Methods 0.000 claims description 2
- 102000023732 binding proteins Human genes 0.000 claims 7
- 108020004414 DNA Proteins 0.000 description 93
- 108010054624 red fluorescent protein Proteins 0.000 description 50
- 238000010362 genome editing Methods 0.000 description 47
- 102100024749 Dynein light chain Tctex-type 1 Human genes 0.000 description 40
- 101000908688 Homo sapiens Dynein light chain Tctex-type 1 Proteins 0.000 description 40
- 102100034051 Heat shock protein HSP 90-alpha Human genes 0.000 description 37
- 101001016865 Homo sapiens Heat shock protein HSP 90-alpha Proteins 0.000 description 37
- 230000000694 effects Effects 0.000 description 33
- 239000002773 nucleotide Substances 0.000 description 31
- 125000003729 nucleotide group Chemical group 0.000 description 31
- 102000014914 Carrier Proteins Human genes 0.000 description 27
- 102100023823 Homeobox protein EMX1 Human genes 0.000 description 22
- 101001048956 Homo sapiens Homeobox protein EMX1 Proteins 0.000 description 22
- 238000013461 design Methods 0.000 description 21
- 101000808011 Homo sapiens Vascular endothelial growth factor A Proteins 0.000 description 20
- 102100039037 Vascular endothelial growth factor A Human genes 0.000 description 20
- 241000588724 Escherichia coli Species 0.000 description 18
- 239000000047 product Substances 0.000 description 17
- 238000003556 assay Methods 0.000 description 16
- 238000002474 experimental method Methods 0.000 description 16
- 238000004458 analytical method Methods 0.000 description 15
- 102000004190 Enzymes Human genes 0.000 description 14
- 108090000790 Enzymes Proteins 0.000 description 14
- 201000010099 disease Diseases 0.000 description 14
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 14
- 102100024364 Disintegrin and metalloproteinase domain-containing protein 8 Human genes 0.000 description 12
- 238000005516 engineering process Methods 0.000 description 12
- 239000012634 fragment Substances 0.000 description 12
- 230000035772 mutation Effects 0.000 description 12
- 102000004196 processed proteins & peptides Human genes 0.000 description 12
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 11
- 241000699670 Mus sp. Species 0.000 description 11
- 230000027455 binding Effects 0.000 description 11
- 238000007481 next generation sequencing Methods 0.000 description 11
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 10
- 101710163270 Nuclease Proteins 0.000 description 10
- 238000003780 insertion Methods 0.000 description 10
- 230000037431 insertion Effects 0.000 description 10
- 239000013612 plasmid Substances 0.000 description 10
- 229920001184 polypeptide Polymers 0.000 description 10
- 238000001890 transfection Methods 0.000 description 10
- 241000588774 Providencia sp. Species 0.000 description 9
- 241000427618 Pseudobacteriovorax antillogorgiicola Species 0.000 description 9
- 230000000875 corresponding effect Effects 0.000 description 9
- 238000012360 testing method Methods 0.000 description 9
- 241001499144 Pantoea brenneri Species 0.000 description 8
- 241000607760 Shigella sonnei Species 0.000 description 8
- 108091028113 Trans-activating crRNA Proteins 0.000 description 8
- 241000543895 Type-F symbiont of Plautia stali Species 0.000 description 8
- 238000005520 cutting process Methods 0.000 description 8
- 230000004927 fusion Effects 0.000 description 8
- 230000004048 modification Effects 0.000 description 8
- 238000012986 modification Methods 0.000 description 8
- 238000007480 sanger sequencing Methods 0.000 description 8
- 229940115939 shigella sonnei Drugs 0.000 description 8
- 241000894007 species Species 0.000 description 8
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 230000008439 repair process Effects 0.000 description 7
- 241000589220 Acetobacter Species 0.000 description 6
- 108700028369 Alleles Proteins 0.000 description 6
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 6
- 238000010453 CRISPR/Cas method Methods 0.000 description 6
- 101001000998 Homo sapiens Protein phosphatase 1 regulatory subunit 12C Proteins 0.000 description 6
- 241000932831 Pantoea stewartii Species 0.000 description 6
- 241000607606 Photobacterium sp. Species 0.000 description 6
- 102100035620 Protein phosphatase 1 regulatory subunit 12C Human genes 0.000 description 6
- 241001022397 Providencia alcalifaciens DSM 30120 Species 0.000 description 6
- 241000588778 Providencia stuartii Species 0.000 description 6
- 241001138501 Salmonella enterica Species 0.000 description 6
- 241000863432 Shewanella putrefaciens Species 0.000 description 6
- 238000000137 annealing Methods 0.000 description 6
- 230000003197 catalytic effect Effects 0.000 description 6
- 238000012217 deletion Methods 0.000 description 6
- 238000009826 distribution Methods 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- 210000004185 liver Anatomy 0.000 description 6
- 210000004072 lung Anatomy 0.000 description 6
- 238000007894 restriction fragment length polymorphism technique Methods 0.000 description 6
- 238000012163 sequencing technique Methods 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 125000006850 spacer group Chemical group 0.000 description 6
- 108091093088 Amplicon Proteins 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 5
- 241000701959 Escherichia virus Lambda Species 0.000 description 5
- 241000124008 Mammalia Species 0.000 description 5
- 108010077850 Nuclear Localization Signals Proteins 0.000 description 5
- 102000002490 Rad51 Recombinase Human genes 0.000 description 5
- 108010068097 Rad51 Recombinase Proteins 0.000 description 5
- 238000013459 approach Methods 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 210000004899 c-terminal region Anatomy 0.000 description 5
- 230000037430 deletion Effects 0.000 description 5
- 239000012091 fetal bovine serum Substances 0.000 description 5
- 238000009396 hybridization Methods 0.000 description 5
- 239000013642 negative control Substances 0.000 description 5
- 230000037361 pathway Effects 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- 241001515965 unidentified phage Species 0.000 description 5
- 102000053602 DNA Human genes 0.000 description 4
- 101100404272 Dictyostelium discoideum redB gene Proteins 0.000 description 4
- 101100316841 Escherichia phage lambda bet gene Proteins 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 108091034117 Oligonucleotide Proteins 0.000 description 4
- 241000611870 Pantoea dispersa Species 0.000 description 4
- 241000804288 Salmonella enterica subsp. enterica serovar Javiana str. 10721 Species 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 4
- 238000007792 addition Methods 0.000 description 4
- 239000011543 agarose gel Substances 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 210000000349 chromosome Anatomy 0.000 description 4
- 230000034431 double-strand break repair via homologous recombination Effects 0.000 description 4
- 239000012636 effector Substances 0.000 description 4
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 4
- 238000012165 high-throughput sequencing Methods 0.000 description 4
- 230000006872 improvement Effects 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 230000001404 mediated effect Effects 0.000 description 4
- 238000011002 quantification Methods 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 239000001509 sodium citrate Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 230000001131 transforming effect Effects 0.000 description 4
- 210000003462 vein Anatomy 0.000 description 4
- 101100272670 Aromatoleum evansii boxB gene Proteins 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 238000010356 CRISPR-Cas9 genome editing Methods 0.000 description 3
- 102100039524 DNA endonuclease RBBP8 Human genes 0.000 description 3
- 108050008316 DNA endonuclease RBBP8 Proteins 0.000 description 3
- 230000033616 DNA repair Effects 0.000 description 3
- 241000701867 Enterobacteria phage T7 Species 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- 208000009869 Neu-Laxova syndrome Diseases 0.000 description 3
- KYRVNWMVYQXFEU-UHFFFAOYSA-N Nocodazole Chemical compound C1=C2NC(NC(=O)OC)=NC2=CC=C1C(=O)C1=CC=CS1 KYRVNWMVYQXFEU-UHFFFAOYSA-N 0.000 description 3
- 241000520272 Pantoea Species 0.000 description 3
- 108091093037 Peptide nucleic acid Proteins 0.000 description 3
- 230000004570 RNA-binding Effects 0.000 description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 3
- 101000910035 Streptococcus pyogenes serotype M1 CRISPR-associated endonuclease Cas9/Csn1 Proteins 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 230000022131 cell cycle Effects 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 3
- 238000012350 deep sequencing Methods 0.000 description 3
- 230000011559 double-strand break repair via nonhomologous end joining Effects 0.000 description 3
- 239000013604 expression vector Substances 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 210000003494 hepatocyte Anatomy 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- -1 morpholino nucleic acid Chemical class 0.000 description 3
- 229950006344 nocodazole Drugs 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 230000000717 retained effect Effects 0.000 description 3
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 3
- 238000007619 statistical method Methods 0.000 description 3
- 230000008685 targeting Effects 0.000 description 3
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 3
- 210000005253 yeast cell Anatomy 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 2
- 229930024421 Adenine Natural products 0.000 description 2
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 2
- 108010011170 Ala-Trp-Arg-His-Pro-Gln-Phe-Gly-Gly Proteins 0.000 description 2
- 101710095342 Apolipoprotein B Proteins 0.000 description 2
- 102100040202 Apolipoprotein B-100 Human genes 0.000 description 2
- 241000203069 Archaea Species 0.000 description 2
- 241000972773 Aulopiformes Species 0.000 description 2
- 208000010061 Autosomal Dominant Polycystic Kidney Diseases 0.000 description 2
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 description 2
- 241000282693 Cercopithecidae Species 0.000 description 2
- 108010079245 Cystic Fibrosis Transmembrane Conductance Regulator Proteins 0.000 description 2
- 102100023419 Cystic fibrosis transmembrane conductance regulator Human genes 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 2
- 230000004568 DNA-binding Effects 0.000 description 2
- 241000702421 Dependoparvovirus Species 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 102100024108 Dystrophin Human genes 0.000 description 2
- 108091029865 Exogenous DNA Proteins 0.000 description 2
- 102000004064 Geminin Human genes 0.000 description 2
- 108090000577 Geminin Proteins 0.000 description 2
- 206010064571 Gene mutation Diseases 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- 101000886596 Homo sapiens Geminin Proteins 0.000 description 2
- 208000026350 Inborn Genetic disease Diseases 0.000 description 2
- 102000000853 LDL receptors Human genes 0.000 description 2
- 108010001831 LDL receptors Proteins 0.000 description 2
- 238000007476 Maximum Likelihood Methods 0.000 description 2
- 108010072388 Methyl-CpG-Binding Protein 2 Proteins 0.000 description 2
- 102100039124 Methyl-CpG-binding protein 2 Human genes 0.000 description 2
- 101000930477 Mus musculus Albumin Proteins 0.000 description 2
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 2
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 2
- 108010052185 Myotonin-Protein Kinase Proteins 0.000 description 2
- 102100022437 Myotonin-protein kinase Human genes 0.000 description 2
- 102100026379 Neurofibromin Human genes 0.000 description 2
- 108010085793 Neurofibromin 1 Proteins 0.000 description 2
- 108091081062 Repeated sequence (DNA) Proteins 0.000 description 2
- 108091028664 Ribonucleotide Proteins 0.000 description 2
- 241000283984 Rodentia Species 0.000 description 2
- 241000607361 Salmonella enterica subsp. enterica Species 0.000 description 2
- 108010003723 Single-Domain Antibodies Proteins 0.000 description 2
- 108020004459 Small interfering RNA Proteins 0.000 description 2
- 238000000692 Student's t-test Methods 0.000 description 2
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 229960000643 adenine Drugs 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 230000000692 anti-sense effect Effects 0.000 description 2
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 230000002759 chromosomal effect Effects 0.000 description 2
- 238000012937 correction Methods 0.000 description 2
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 239000005547 deoxyribonucleotide Substances 0.000 description 2
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 229960000633 dextran sulfate Drugs 0.000 description 2
- 238000004520 electroporation Methods 0.000 description 2
- 210000001671 embryonic stem cell Anatomy 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 238000003197 gene knockdown Methods 0.000 description 2
- 208000016361 genetic disease Diseases 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 2
- 235000003642 hunger Nutrition 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 230000004807 localization Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 108091070501 miRNA Proteins 0.000 description 2
- 239000002679 microRNA Substances 0.000 description 2
- 108091027963 non-coding RNA Proteins 0.000 description 2
- 102000042567 non-coding RNA Human genes 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 230000009438 off-target cleavage Effects 0.000 description 2
- 230000009437 off-target effect Effects 0.000 description 2
- 201000008519 polycystic kidney disease 1 Diseases 0.000 description 2
- 201000008542 polycystic kidney disease 2 Diseases 0.000 description 2
- 108700032676 polycystic kidney disease 2 Proteins 0.000 description 2
- 230000003234 polygenic effect Effects 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 210000001236 prokaryotic cell Anatomy 0.000 description 2
- 239000012264 purified product Substances 0.000 description 2
- 230000022983 regulation of cell cycle Effects 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000002336 ribonucleotide Substances 0.000 description 2
- 125000002652 ribonucleotide group Chemical group 0.000 description 2
- 235000019515 salmon Nutrition 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000004055 small Interfering RNA Substances 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000001488 sodium phosphate Substances 0.000 description 2
- 229910000162 sodium phosphate Inorganic materials 0.000 description 2
- 230000037351 starvation Effects 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 238000012353 t test Methods 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 229940104230 thymidine Drugs 0.000 description 2
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- QIVUCLWGARAQIO-OLIXTKCUSA-N (3s)-n-[(3s,5s,6r)-6-methyl-2-oxo-1-(2,2,2-trifluoroethyl)-5-(2,3,6-trifluorophenyl)piperidin-3-yl]-2-oxospiro[1h-pyrrolo[2,3-b]pyridine-3,6'-5,7-dihydrocyclopenta[b]pyridine]-3'-carboxamide Chemical compound C1([C@H]2[C@H](N(C(=O)[C@@H](NC(=O)C=3C=C4C[C@]5(CC4=NC=3)C3=CC=CN=C3NC5=O)C2)CC(F)(F)F)C)=C(F)C=CC(F)=C1F QIVUCLWGARAQIO-OLIXTKCUSA-N 0.000 description 1
- 230000006269 (delayed) early viral mRNA transcription Effects 0.000 description 1
- 101710169336 5'-deoxyadenosine deaminase Proteins 0.000 description 1
- 241000093740 Acidaminococcus sp. Species 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 102000055025 Adenosine deaminases Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 101710081722 Antitrypsin Proteins 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 102000002804 Ataxia Telangiectasia Mutated Proteins Human genes 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 208000020925 Bipolar disease Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000193764 Brevibacillus brevis Species 0.000 description 1
- 108091079001 CRISPR RNA Proteins 0.000 description 1
- 238000010354 CRISPR gene editing Methods 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 102000053642 Catalytic RNA Human genes 0.000 description 1
- 108090000994 Catalytic RNA Proteins 0.000 description 1
- 241000700198 Cavia Species 0.000 description 1
- 229940123587 Cell cycle inhibitor Drugs 0.000 description 1
- 101100007328 Cocos nucifera COS-1 gene Proteins 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 208000002330 Congenital Heart Defects Diseases 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- 102000011724 DNA Repair Enzymes Human genes 0.000 description 1
- 108010076525 DNA Repair Enzymes Proteins 0.000 description 1
- 238000007702 DNA assembly Methods 0.000 description 1
- 238000010442 DNA editing Methods 0.000 description 1
- 102000003844 DNA helicases Human genes 0.000 description 1
- 108090000133 DNA helicases Proteins 0.000 description 1
- 102000052510 DNA-Binding Proteins Human genes 0.000 description 1
- 101710096438 DNA-binding protein Proteins 0.000 description 1
- 108010069091 Dystrophin Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 102000004533 Endonucleases Human genes 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 108010059378 Endopeptidases Proteins 0.000 description 1
- 102000005593 Endopeptidases Human genes 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 241000588722 Escherichia Species 0.000 description 1
- 102000001690 Factor VIII Human genes 0.000 description 1
- 108010054218 Factor VIII Proteins 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 229920001917 Ficoll Polymers 0.000 description 1
- 108091092584 GDNA Proteins 0.000 description 1
- 102100039956 Geminin Human genes 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 108091027305 Heteroduplex Proteins 0.000 description 1
- 241001640034 Heteropterys Species 0.000 description 1
- 102100029054 Homeobox protein notochord Human genes 0.000 description 1
- 241001272567 Hominoidea Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000634521 Homo sapiens Homeobox protein notochord Proteins 0.000 description 1
- 101001006794 Homo sapiens Kinesin-like protein KIF6 Proteins 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 102100027927 Kinesin-like protein KIF6 Human genes 0.000 description 1
- 241000235649 Kluyveromyces Species 0.000 description 1
- 101150105104 Kras gene Proteins 0.000 description 1
- 241000904817 Lachnospiraceae bacterium Species 0.000 description 1
- 208000024556 Mendelian disease Diseases 0.000 description 1
- YBHQCJILTOVLHD-YVMONPNESA-N Mirin Chemical compound S1C(N)=NC(=O)\C1=C\C1=CC=C(O)C=C1 YBHQCJILTOVLHD-YVMONPNESA-N 0.000 description 1
- 206010029260 Neuroblastoma Diseases 0.000 description 1
- 101100462611 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) prr-1 gene Proteins 0.000 description 1
- 229940122426 Nuclease inhibitor Drugs 0.000 description 1
- 108091005461 Nucleic proteins Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 238000010222 PCR analysis Methods 0.000 description 1
- 241000282579 Pan Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 241000235648 Pichia Species 0.000 description 1
- 241001527110 Plautia Species 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 241000709748 Pseudomonas phage PRR1 Species 0.000 description 1
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 1
- 102000009572 RNA Polymerase II Human genes 0.000 description 1
- 108010009460 RNA Polymerase II Proteins 0.000 description 1
- 102000044126 RNA-Binding Proteins Human genes 0.000 description 1
- 108700020471 RNA-Binding Proteins Proteins 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 241000293825 Rhinosporidium Species 0.000 description 1
- 101710170912 SOSS complex subunit B1 Proteins 0.000 description 1
- 241000235070 Saccharomyces Species 0.000 description 1
- 101100068077 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) GCN2 gene Proteins 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 241000235346 Schizosaccharomyces Species 0.000 description 1
- 241000607768 Shigella Species 0.000 description 1
- 101710141933 Single-stranded DNA-binding protein 1 Proteins 0.000 description 1
- 102100029719 Single-stranded DNA-binding protein, mitochondrial Human genes 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 108091027544 Subgenomic mRNA Proteins 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 108700019146 Transgenes Proteins 0.000 description 1
- 102000018390 Ubiquitin-Specific Proteases Human genes 0.000 description 1
- 108010066496 Ubiquitin-Specific Proteases Proteins 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 210000005006 adaptive immune system Anatomy 0.000 description 1
- 239000012082 adaptor molecule Substances 0.000 description 1
- 102000035181 adaptor proteins Human genes 0.000 description 1
- 108091005764 adaptor proteins Proteins 0.000 description 1
- 101150063416 add gene Proteins 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000001475 anti-trypsic effect Effects 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 230000002457 bidirectional effect Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 210000004671 cell-free system Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000013000 chemical inhibitor Substances 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 208000016653 cleft lip/palate Diseases 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000010225 co-occurrence analysis Methods 0.000 description 1
- 229940105778 coagulation factor viii Drugs 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000009918 complex formation Effects 0.000 description 1
- 238000004590 computer program Methods 0.000 description 1
- 208000028831 congenital heart disease Diseases 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 229940104302 cytosine Drugs 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000009547 development abnormality Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 210000001840 diploid cell Anatomy 0.000 description 1
- 230000005782 double-strand break Effects 0.000 description 1
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 206010015037 epilepsy Diseases 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 235000019688 fish Nutrition 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000007614 genetic variation Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 102000054910 human GMNN Human genes 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000021005 inheritance pattern Diseases 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 108010082117 matrigel Proteins 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000002493 microarray Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 230000011278 mitosis Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 201000010193 neural tube defect Diseases 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 201000007909 oculocutaneous albinism Diseases 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 235000015927 pasta Nutrition 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000012660 pharmacological inhibitor Substances 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 208000030683 polygenic disease Diseases 0.000 description 1
- 230000004481 post-translational protein modification Effects 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000010469 pro-virus integration Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000031877 prophase Effects 0.000 description 1
- 230000012743 protein tagging Effects 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 108091092562 ribozyme Proteins 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 238000001963 scanning near-field photolithography Methods 0.000 description 1
- 201000000980 schizophrenia Diseases 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 125000003607 serino group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C(O[H])([H])[H] 0.000 description 1
- 230000003007 single stranded DNA break Effects 0.000 description 1
- FQENQNTWSFEDLI-UHFFFAOYSA-J sodium diphosphate Chemical compound [Na+].[Na+].[Na+].[Na+].[O-]P([O-])(=O)OP([O-])([O-])=O FQENQNTWSFEDLI-UHFFFAOYSA-J 0.000 description 1
- 239000012064 sodium phosphate buffer Substances 0.000 description 1
- 229940048086 sodium pyrophosphate Drugs 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 208000035581 susceptibility to neural tube defects Diseases 0.000 description 1
- 230000001360 synchronised effect Effects 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 235000019818 tetrasodium diphosphate Nutrition 0.000 description 1
- 239000001577 tetrasodium phosphonato phosphate Substances 0.000 description 1
- 229940113082 thymine Drugs 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
- 239000002753 trypsin inhibitor Substances 0.000 description 1
- 230000005740 tumor formation Effects 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 108700026220 vif Genes Proteins 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/102—Mutagenizing nucleic acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/24—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- C07K14/245—Escherichia (G)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/305—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F)
- C07K14/31—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F) from Staphylococcus (G)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/115—Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
- C12N15/625—DNA sequences coding for fusion proteins containing a sequence coding for a signal sequence
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/90—Stable introduction of foreign DNA into chromosome
- C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
- C12N15/907—Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/22—Ribonucleases [RNase]; Deoxyribonucleases [DNase]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/09—Fusion polypeptide containing a localisation/targetting motif containing a nuclear localisation signal
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/16—Aptamers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/20—Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPR]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/35—Nature of the modification
- C12N2310/351—Conjugate
- C12N2310/3519—Fusion with another nucleic acid
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2510/00—Genetically modified cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2800/00—Nucleic acids vectors
- C12N2800/80—Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Crystallography & Structural Chemistry (AREA)
- Cell Biology (AREA)
- Mycology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
- Saccharide Compounds (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US202062984618P | 2020-03-03 | 2020-03-03 | |
| US202163146447P | 2021-02-05 | 2021-02-05 | |
| PCT/US2021/020513 WO2021178432A1 (en) | 2020-03-03 | 2021-03-02 | Rna-guided genome recombineering at kilobase scale |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| IL296057A true IL296057A (en) | 2022-10-01 |
Family
ID=77614129
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| IL296057A IL296057A (en) | 2020-03-03 | 2021-03-02 | Ribonucleic acid-guided genome recombination engineering at the scale of thousands of bases |
Country Status (11)
| Country | Link |
|---|---|
| US (1) | US20230091242A1 (https=) |
| EP (1) | EP4114845A4 (https=) |
| JP (1) | JP2023515670A (https=) |
| KR (1) | KR20220151175A (https=) |
| CN (1) | CN115667283A (https=) |
| AU (1) | AU2021231769A1 (https=) |
| BR (1) | BR112022017196A2 (https=) |
| CA (1) | CA3173526A1 (https=) |
| IL (1) | IL296057A (https=) |
| MX (1) | MX2022010835A (https=) |
| WO (1) | WO2021178432A1 (https=) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4396340A4 (en) * | 2021-09-01 | 2025-09-24 | Univ Leland Stanford Junior | RNA-GUIDED GENOME RECOMBINATION AT THE KILOBASE SCALE |
| JP2025505732A (ja) * | 2022-02-10 | 2025-02-28 | ザ ボード オブ トラスティーズ オブ ザ レランド スタンフォード ジュニア ユニバーシティー | キロ塩基スケールでのｒｎａガイドゲノムリコンビニアリング |
| WO2023154892A1 (en) * | 2022-02-10 | 2023-08-17 | Possible Medicines Llc | Rna-guided genome recombineering at kilobase scale |
| WO2024168265A1 (en) * | 2023-02-10 | 2024-08-15 | Possible Medicines Llc | Aav delivery of rna guided recombination system |
| WO2024168253A1 (en) * | 2023-02-10 | 2024-08-15 | Possible Medicines Llc | Delivery of an rna guided recombination system |
| CN120555441B (zh) * | 2025-05-29 | 2026-04-17 | 西北农林科技大学 | 基于CtIP蛋白提高CRISPR/Cas9介导的外源基因整合效率的DNA适配体及其应用 |
Family Cites Families (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU752105B2 (en) * | 1997-12-05 | 2002-09-05 | Europaisches Laboratorium Fur Molekularbiologie (Embl) | Novel dna cloning method |
| US9388430B2 (en) * | 2013-09-06 | 2016-07-12 | President And Fellows Of Harvard College | Cas9-recombinase fusion proteins and uses thereof |
| WO2016022363A2 (en) * | 2014-07-30 | 2016-02-11 | President And Fellows Of Harvard College | Cas9 proteins including ligand-dependent inteins |
| EP3265559B1 (en) * | 2015-03-03 | 2021-01-06 | The General Hospital Corporation | Engineered crispr-cas9 nucleases with altered pam specificity |
| WO2016205759A1 (en) * | 2015-06-18 | 2016-12-22 | The Broad Institute Inc. | Engineering and optimization of systems, methods, enzymes and guide scaffolds of cas9 orthologs and variants for sequence manipulation |
| US11479793B2 (en) * | 2015-07-15 | 2022-10-25 | Rutgers, The State University Of New Jersey | Nuclease-independent targeted gene editing platform and uses thereof |
| WO2019089910A1 (en) * | 2017-11-01 | 2019-05-09 | Ohio State Innovation Foundation | Highly compact cas9-based transcriptional regulators for in vivo gene regulation |
| KR20210152597A (ko) * | 2017-12-22 | 2021-12-15 | (주)지플러스생명과학 | 키메라 게놈 조작 분자 및 방법 |
| EP4396340A4 (en) * | 2021-09-01 | 2025-09-24 | Univ Leland Stanford Junior | RNA-GUIDED GENOME RECOMBINATION AT THE KILOBASE SCALE |
-
2021
- 2021-03-02 IL IL296057A patent/IL296057A/en unknown
- 2021-03-02 CA CA3173526A patent/CA3173526A1/en active Pending
- 2021-03-02 EP EP21764351.9A patent/EP4114845A4/en active Pending
- 2021-03-02 WO PCT/US2021/020513 patent/WO2021178432A1/en not_active Ceased
- 2021-03-02 KR KR1020227033540A patent/KR20220151175A/ko active Pending
- 2021-03-02 AU AU2021231769A patent/AU2021231769A1/en active Pending
- 2021-03-02 BR BR112022017196A patent/BR112022017196A2/pt unknown
- 2021-03-02 MX MX2022010835A patent/MX2022010835A/es unknown
- 2021-03-02 JP JP2022552549A patent/JP2023515670A/ja active Pending
- 2021-03-02 US US17/905,457 patent/US20230091242A1/en active Pending
- 2021-03-02 CN CN202180033011.8A patent/CN115667283A/zh active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| EP4114845A1 (en) | 2023-01-11 |
| JP2023515670A (ja) | 2023-04-13 |
| MX2022010835A (es) | 2022-09-29 |
| AU2021231769A1 (en) | 2022-09-29 |
| WO2021178432A1 (en) | 2021-09-10 |
| KR20220151175A (ko) | 2022-11-14 |
| EP4114845A4 (en) | 2024-03-06 |
| CN115667283A (zh) | 2023-01-31 |
| US20230091242A1 (en) | 2023-03-23 |
| WO2021178432A9 (en) | 2021-10-28 |
| BR112022017196A2 (pt) | 2022-10-25 |
| CA3173526A1 (en) | 2021-09-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| IL296057A (en) | Ribonucleic acid-guided genome recombination engineering at the scale of thousands of bases | |
| JP7605852B2 (ja) | クラスｉｉのｖ型ｃｒｉｓｐｒ系 | |
| JP7705382B2 (ja) | 新規ｃｒｉｓｐｒ ｄｎａターゲティング酵素及びシステム | |
| US20200407716A1 (en) | Novel crispr dna targeting enzymes and systems | |
| Mosberg et al. | Improving lambda red genome engineering in Escherichia coli via rational removal of endogenous nucleases | |
| CA3192224A1 (en) | Base editing enzymes | |
| KR20240055073A (ko) | 클래스 ii, v형 crispr 시스템 | |
| EP4567117A2 (en) | Engineered mad7 directed endonuclease | |
| CN111315890A (zh) | 提高同源重组的方法和其组合物 | |
| CA3234233A1 (en) | Endonuclease systems | |
| WO2019120193A1 (zh) | 拆分型单碱基基因编辑系统及其应用 | |
| CN116376945B (zh) | 一种I型Casposase基因插入工具及应用 | |
| JP2024501892A (ja) | 新規の核酸誘導型ヌクレアーゼ | |
| US11332751B2 (en) | Methods and compositions for production of aromatic and other compounds in yeast | |
| IL286917B (en) | Methods for scar-free insertion of targeted modifications into targeted vectors | |
| CN105087554B (zh) | Dna磷硫酰化修饰基因簇 | |
| Guo et al. | A Novel Double-Stranded DNA Deaminase-Based and Transcriptional Activator-Assisted Nuclear and Mitochondrial Cytosine Base Editors with Expanded Target Compatibility and Enhanced Activity | |
| US20250059568A1 (en) | Class ii, type v crispr systems | |
| Sung et al. | Scarless chromosomal gene knockout methods | |
| CN104357422A (zh) | 转录激活子样效应因子核酸酶及其编码基因与应用 | |
| JP2025530183A (ja) | レット症候群治療法 | |
| CN117417916A (zh) | 酪氨酸整合酶、核酸分子、载体、宿主细胞、应用、遗传操作工具 | |
| HK40065084A (en) | Enzymes with ruvc domains | |
| WO2019237389A1 (zh) | CRISPR/Cas9靶向敲除人MTABC3基因及其特异性gRNA |