CA2936646C - Methods and compositions for sequences guiding cas9 targeting - Google Patents
Methods and compositions for sequences guiding cas9 targeting Download PDFInfo
- Publication number
- CA2936646C CA2936646C CA2936646A CA2936646A CA2936646C CA 2936646 C CA2936646 C CA 2936646C CA 2936646 A CA2936646 A CA 2936646A CA 2936646 A CA2936646 A CA 2936646A CA 2936646 C CA2936646 C CA 2936646C
- Authority
- CA
- Canada
- Prior art keywords
- sequence
- nucleic acid
- acid construct
- nucleotides
- stitch
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 230000008685 targeting Effects 0.000 title claims abstract description 31
- 238000000034 method Methods 0.000 title abstract description 37
- 239000000203 mixture Substances 0.000 title abstract description 11
- 101150038500 cas9 gene Proteins 0.000 title description 4
- 239000002773 nucleotide Substances 0.000 claims description 470
- 125000003729 nucleotide group Chemical group 0.000 claims description 466
- 150000007523 nucleic acids Chemical class 0.000 claims description 378
- 102000039446 nucleic acids Human genes 0.000 claims description 373
- 108020004707 nucleic acids Proteins 0.000 claims description 373
- 108091033409 CRISPR Proteins 0.000 claims description 206
- 108020004414 DNA Proteins 0.000 claims description 187
- 238000011144 upstream manufacturing Methods 0.000 claims description 145
- 101710163270 Nuclease Proteins 0.000 claims description 118
- 125000006850 spacer group Chemical group 0.000 claims description 105
- 230000000295 complement effect Effects 0.000 claims description 94
- 210000004027 cell Anatomy 0.000 claims description 89
- 230000000694 effects Effects 0.000 claims description 74
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 60
- 229920001184 polypeptide Polymers 0.000 claims description 59
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 58
- 238000003776 cleavage reaction Methods 0.000 claims description 47
- 230000007017 scission Effects 0.000 claims description 47
- 230000014509 gene expression Effects 0.000 claims description 41
- 150000001413 amino acids Chemical group 0.000 claims description 35
- 238000009396 hybridization Methods 0.000 claims description 33
- 230000035772 mutation Effects 0.000 claims description 31
- 241000282414 Homo sapiens Species 0.000 claims description 15
- 230000004048 modification Effects 0.000 claims description 13
- 238000012986 modification Methods 0.000 claims description 13
- 241000196324 Embryophyta Species 0.000 claims description 11
- 241000894006 Bacteria Species 0.000 claims description 10
- 108020004705 Codon Proteins 0.000 claims description 9
- 102000053602 DNA Human genes 0.000 claims description 6
- 241000238631 Hexapoda Species 0.000 claims description 6
- 241000251468 Actinopterygii Species 0.000 claims description 5
- 241000243321 Cnidaria Species 0.000 claims description 5
- 241000186606 Lactobacillus gasseri Species 0.000 claims description 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 5
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 5
- 241000617156 archaeon Species 0.000 claims description 5
- 241000244203 Caenorhabditis elegans Species 0.000 claims description 4
- 241000255601 Drosophila melanogaster Species 0.000 claims description 4
- 244000068988 Glycine max Species 0.000 claims description 4
- 235000010469 Glycine max Nutrition 0.000 claims description 4
- 244000299507 Gossypium hirsutum Species 0.000 claims description 4
- 235000009432 Gossypium hirsutum Nutrition 0.000 claims description 4
- 101000615488 Homo sapiens Methyl-CpG-binding domain protein 2 Proteins 0.000 claims description 4
- 241000124008 Mammalia Species 0.000 claims description 4
- 102100021299 Methyl-CpG-binding domain protein 2 Human genes 0.000 claims description 4
- 108060004795 Methyltransferase Proteins 0.000 claims description 4
- 241000700157 Rattus norvegicus Species 0.000 claims description 4
- 241000235070 Saccharomyces Species 0.000 claims description 4
- 102000006275 Ubiquitin-Protein Ligases Human genes 0.000 claims description 4
- 108010083111 Ubiquitin-Protein Ligases Proteins 0.000 claims description 4
- 102000005421 acetyltransferase Human genes 0.000 claims description 4
- 108020002494 acetyltransferase Proteins 0.000 claims description 4
- 230000030648 nucleus localization Effects 0.000 claims description 4
- 230000008439 repair process Effects 0.000 claims description 4
- 108091035539 telomere Proteins 0.000 claims description 4
- 102000055501 telomere Human genes 0.000 claims description 4
- 210000003411 telomere Anatomy 0.000 claims description 4
- 241000219195 Arabidopsis thaliana Species 0.000 claims description 3
- 230000005778 DNA damage Effects 0.000 claims description 3
- 231100000277 DNA damage Toxicity 0.000 claims description 3
- 230000033616 DNA repair Effects 0.000 claims description 3
- 108010046331 Deoxyribodipyrimidine photo-lyase Proteins 0.000 claims description 3
- 241000233866 Fungi Species 0.000 claims description 3
- 102100034343 Integrase Human genes 0.000 claims description 3
- 108010061833 Integrases Proteins 0.000 claims description 3
- 102000003960 Ligases Human genes 0.000 claims description 3
- 108090000364 Ligases Proteins 0.000 claims description 3
- 241001465754 Metazoa Species 0.000 claims description 3
- 241000699660 Mus musculus Species 0.000 claims description 3
- 230000010718 Oxidation Activity Effects 0.000 claims description 3
- 102000045595 Phosphoprotein Phosphatases Human genes 0.000 claims description 3
- 108700019535 Phosphoprotein Phosphatases Proteins 0.000 claims description 3
- 102000001253 Protein Kinase Human genes 0.000 claims description 3
- 102000055027 Protein Methyltransferases Human genes 0.000 claims description 3
- 108700040121 Protein Methyltransferases Proteins 0.000 claims description 3
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 claims description 3
- 108091093078 Pyrimidine dimer Proteins 0.000 claims description 3
- 102000018120 Recombinases Human genes 0.000 claims description 3
- 108010091086 Recombinases Proteins 0.000 claims description 3
- 108010017842 Telomerase Proteins 0.000 claims description 3
- 102000008579 Transposases Human genes 0.000 claims description 3
- 108010020764 Transposases Proteins 0.000 claims description 3
- 230000006154 adenylylation Effects 0.000 claims description 3
- 230000029936 alkylation Effects 0.000 claims description 3
- 238000005804 alkylation reaction Methods 0.000 claims description 3
- 230000001580 bacterial effect Effects 0.000 claims description 3
- 230000009615 deamination Effects 0.000 claims description 3
- 238000006481 deamination reaction Methods 0.000 claims description 3
- 230000006114 demyristoylation Effects 0.000 claims description 3
- 230000027832 depurination Effects 0.000 claims description 3
- 230000002538 fungal effect Effects 0.000 claims description 3
- 230000007498 myristoylation Effects 0.000 claims description 3
- 108060006633 protein kinase Proteins 0.000 claims description 3
- 239000013635 pyrimidine dimer Substances 0.000 claims description 3
- 210000004102 animal cell Anatomy 0.000 claims description 2
- 210000004602 germ cell Anatomy 0.000 claims description 2
- 210000005260 human cell Anatomy 0.000 claims description 2
- 210000004962 mammalian cell Anatomy 0.000 claims description 2
- 210000001082 somatic cell Anatomy 0.000 claims description 2
- 210000000130 stem cell Anatomy 0.000 claims description 2
- 238000000968 medical method and process Methods 0.000 claims 9
- VUFNLQXQSDUXKB-DOFZRALJSA-N 2-[4-[4-[bis(2-chloroethyl)amino]phenyl]butanoyloxy]ethyl (5z,8z,11z,14z)-icosa-5,8,11,14-tetraenoate Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(=O)OCCOC(=O)CCCC1=CC=C(N(CCCl)CCCl)C=C1 VUFNLQXQSDUXKB-DOFZRALJSA-N 0.000 claims 3
- 108090000623 proteins and genes Proteins 0.000 abstract description 22
- 102000004169 proteins and genes Human genes 0.000 abstract description 13
- 238000010362 genome editing Methods 0.000 abstract description 5
- 108091028043 Nucleic acid sequence Proteins 0.000 description 42
- 239000013612 plasmid Substances 0.000 description 37
- 102000040430 polynucleotide Human genes 0.000 description 36
- 108091033319 polynucleotide Proteins 0.000 description 36
- 239000002157 polynucleotide Substances 0.000 description 36
- 230000009466 transformation Effects 0.000 description 34
- 108091035707 Consensus sequence Proteins 0.000 description 24
- 108091028113 Trans-activating crRNA Proteins 0.000 description 22
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 21
- 238000003499 nucleic acid array Methods 0.000 description 21
- 230000027455 binding Effects 0.000 description 16
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 15
- 238000012360 testing method Methods 0.000 description 15
- 239000013598 vector Substances 0.000 description 15
- 241000194020 Streptococcus thermophilus Species 0.000 description 12
- 238000003556 assay Methods 0.000 description 12
- 230000006870 function Effects 0.000 description 10
- 238000002887 multiple sequence alignment Methods 0.000 description 10
- 241000894007 species Species 0.000 description 10
- 239000002609 medium Substances 0.000 description 9
- 230000001105 regulatory effect Effects 0.000 description 9
- 108091079001 CRISPR RNA Proteins 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 8
- 230000002068 genetic effect Effects 0.000 description 8
- 108091026890 Coding region Proteins 0.000 description 7
- 108020005004 Guide RNA Proteins 0.000 description 7
- 241000186660 Lactobacillus Species 0.000 description 7
- 108091027544 Subgenomic mRNA Proteins 0.000 description 7
- 230000001965 increasing effect Effects 0.000 description 7
- 239000003550 marker Substances 0.000 description 7
- 108020004635 Complementary DNA Proteins 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 238000007476 Maximum Likelihood Methods 0.000 description 6
- 108700019146 Transgenes Proteins 0.000 description 6
- 101150063416 add gene Proteins 0.000 description 6
- 239000002299 complementary DNA Substances 0.000 description 6
- 229940039696 lactobacillus Drugs 0.000 description 6
- -1 tracrRNA Chemical class 0.000 description 6
- 108700010070 Codon Usage Proteins 0.000 description 5
- 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 5
- 241000218588 Lactobacillus rhamnosus Species 0.000 description 5
- 241000193996 Streptococcus pyogenes Species 0.000 description 5
- 230000001186 cumulative effect Effects 0.000 description 5
- 238000003752 polymerase chain reaction Methods 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 238000002864 sequence alignment Methods 0.000 description 5
- 238000013518 transcription Methods 0.000 description 5
- 230000035897 transcription Effects 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- 108010042407 Endonucleases Proteins 0.000 description 4
- 102000004533 Endonucleases Human genes 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 4
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 4
- 238000012300 Sequence Analysis Methods 0.000 description 4
- 241000194017 Streptococcus Species 0.000 description 4
- 238000010804 cDNA synthesis Methods 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 108020004999 messenger RNA Proteins 0.000 description 4
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 3
- 238000010354 CRISPR gene editing Methods 0.000 description 3
- 238000010446 CRISPR interference Methods 0.000 description 3
- 241000186679 Lactobacillus buchneri Species 0.000 description 3
- 241000752990 Lactobacillus buchneri CD034 Species 0.000 description 3
- 244000199866 Lactobacillus casei Species 0.000 description 3
- 235000013958 Lactobacillus casei Nutrition 0.000 description 3
- 102000003661 Ribonuclease III Human genes 0.000 description 3
- 108010057163 Ribonuclease III Proteins 0.000 description 3
- 230000004721 adaptive immunity Effects 0.000 description 3
- 238000007792 addition Methods 0.000 description 3
- 238000009395 breeding Methods 0.000 description 3
- 230000001488 breeding effect Effects 0.000 description 3
- 210000000349 chromosome Anatomy 0.000 description 3
- 230000005782 double-strand break Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 230000000977 initiatory effect Effects 0.000 description 3
- 230000010354 integration Effects 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 229940017800 lactobacillus casei Drugs 0.000 description 3
- 238000005457 optimization Methods 0.000 description 3
- 229920002401 polyacrylamide Polymers 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 230000002103 transcriptional effect Effects 0.000 description 3
- 230000001052 transient effect Effects 0.000 description 3
- FVFVNNKYKYZTJU-UHFFFAOYSA-N 6-chloro-1,3,5-triazine-2,4-diamine Chemical compound NC1=NC(N)=NC(Cl)=N1 FVFVNNKYKYZTJU-UHFFFAOYSA-N 0.000 description 2
- 108020005544 Antisense RNA Proteins 0.000 description 2
- 241000203069 Archaea Species 0.000 description 2
- 230000007018 DNA scission Effects 0.000 description 2
- 230000004568 DNA-binding Effects 0.000 description 2
- 108010008532 Deoxyribonuclease I Proteins 0.000 description 2
- 102000007260 Deoxyribonuclease I Human genes 0.000 description 2
- 241001646716 Escherichia coli K-12 Species 0.000 description 2
- 241000186684 Lactobacillus pentosus Species 0.000 description 2
- 241000272168 Laridae Species 0.000 description 2
- 238000000636 Northern blotting Methods 0.000 description 2
- 108020004711 Nucleic Acid Probes Proteins 0.000 description 2
- 108091005461 Nucleic proteins Proteins 0.000 description 2
- 108091081062 Repeated sequence (DNA) Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000002105 Southern blotting Methods 0.000 description 2
- 241001633172 Streptococcus thermophilus LMD-9 Species 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 230000000692 anti-sense effect Effects 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 210000003763 chloroplast Anatomy 0.000 description 2
- 239000003184 complementary RNA Substances 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 229920000140 heteropolymer Polymers 0.000 description 2
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 239000002679 microRNA Substances 0.000 description 2
- 238000010369 molecular cloning Methods 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000002853 nucleic acid probe Substances 0.000 description 2
- 230000008520 organization Effects 0.000 description 2
- 238000004806 packaging method and process Methods 0.000 description 2
- 210000002706 plastid Anatomy 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 229910001415 sodium ion Inorganic materials 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 241001515965 unidentified phage Species 0.000 description 2
- 108010052418 (N-(2-((4-((2-((4-(9-acridinylamino)phenyl)amino)-2-oxoethyl)amino)-4-oxobutyl)amino)-1-(1H-imidazol-4-ylmethyl)-1-oxoethyl)-6-(((-2-aminoethyl)amino)methyl)-2-pyridinecarboxamidato) iron(1+) Proteins 0.000 description 1
- UHDGCWIWMRVCDJ-UHFFFAOYSA-N 1-beta-D-Xylofuranosyl-NH-Cytosine Natural products O=C1N=C(N)C=CN1C1C(O)C(O)C(CO)O1 UHDGCWIWMRVCDJ-UHFFFAOYSA-N 0.000 description 1
- LRSASMSXMSNRBT-UHFFFAOYSA-N 5-methylcytosine Chemical compound CC1=CNC(=O)N=C1N LRSASMSXMSNRBT-UHFFFAOYSA-N 0.000 description 1
- CKOMXBHMKXXTNW-UHFFFAOYSA-N 6-methyladenine Chemical compound CNC1=NC=NC2=C1N=CN2 CKOMXBHMKXXTNW-UHFFFAOYSA-N 0.000 description 1
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 241000186045 Actinomyces naeslundii Species 0.000 description 1
- 241000589158 Agrobacterium Species 0.000 description 1
- 108020003566 Antisense Oligodeoxyribonucleotides Proteins 0.000 description 1
- 241000219194 Arabidopsis Species 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 102000053642 Catalytic RNA Human genes 0.000 description 1
- 108090000994 Catalytic RNA Proteins 0.000 description 1
- UHDGCWIWMRVCDJ-PSQAKQOGSA-N Cytidine Natural products O=C1N=C(N)C=CN1[C@@H]1[C@@H](O)[C@@H](O)[C@H](CO)O1 UHDGCWIWMRVCDJ-PSQAKQOGSA-N 0.000 description 1
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 description 1
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 1
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 1
- 241000702421 Dependoparvovirus Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000975394 Evechinus chloroticus Species 0.000 description 1
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 description 1
- 229930010555 Inosine Natural products 0.000 description 1
- UGQMRVRMYYASKQ-KQYNXXCUSA-N Inosine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C2=NC=NC(O)=C2N=C1 UGQMRVRMYYASKQ-KQYNXXCUSA-N 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 241000186712 Lactobacillus animalis Species 0.000 description 1
- 241000762071 Lactobacillus curvatus CRL 705 Species 0.000 description 1
- 241000186841 Lactobacillus farciminis Species 0.000 description 1
- 241001561398 Lactobacillus jensenii Species 0.000 description 1
- 241000917009 Lactobacillus rhamnosus GG Species 0.000 description 1
- 239000006142 Luria-Bertani Agar Substances 0.000 description 1
- 102000016397 Methyltransferase Human genes 0.000 description 1
- 108010085220 Multiprotein Complexes Proteins 0.000 description 1
- 102000007474 Multiprotein Complexes Human genes 0.000 description 1
- 108020004485 Nonsense Codon Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 1
- 108091006629 SLC13A2 Proteins 0.000 description 1
- 108700026226 TATA Box Proteins 0.000 description 1
- DRTQHJPVMGBUCF-XVFCMESISA-N Uridine Natural products O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-XVFCMESISA-N 0.000 description 1
- 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 Chemical class 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 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 230000003466 anti-cipated effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000003293 antisense oligodeoxyribonucleotide Substances 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 238000003491 array Methods 0.000 description 1
- 210000004507 artificial chromosome Anatomy 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- DRTQHJPVMGBUCF-PSQAKQOGSA-N beta-L-uridine Natural products O[C@H]1[C@@H](O)[C@H](CO)O[C@@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-PSQAKQOGSA-N 0.000 description 1
- 230000003851 biochemical process Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000008436 biogenesis Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 108091036078 conserved sequence Proteins 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000013601 cosmid vector Substances 0.000 description 1
- UHDGCWIWMRVCDJ-ZAKLUEHWSA-N cytidine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O1 UHDGCWIWMRVCDJ-ZAKLUEHWSA-N 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000000368 destabilizing effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000000635 electron micrograph Methods 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 231100000221 frame shift mutation induction Toxicity 0.000 description 1
- 230000037433 frameshift Effects 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000015788 innate immune response Effects 0.000 description 1
- 229960003786 inosine Drugs 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000011005 laboratory method Methods 0.000 description 1
- 229940059406 lactobacillus rhamnosus gg Drugs 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000007726 management method Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 108091070501 miRNA Proteins 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 238000007899 nucleic acid hybridization Methods 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 230000009438 off-target cleavage Effects 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 150000004713 phosphodiesters Chemical group 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000005912 ran GTP Binding Protein Human genes 0.000 description 1
- 230000008707 rearrangement Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000022532 regulation of transcription, DNA-dependent Effects 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 108091092562 ribozyme Proteins 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000013605 shuttle vector Substances 0.000 description 1
- 230000003007 single stranded DNA break Effects 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000002123 temporal effect Effects 0.000 description 1
- OFVLGDICTFRJMM-WESIUVDSSA-N tetracycline Chemical compound C1=CC=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O OFVLGDICTFRJMM-WESIUVDSSA-N 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 125000000430 tryptophan group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C2=C([H])C([H])=C([H])C([H])=C12 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- DRTQHJPVMGBUCF-UHFFFAOYSA-N uracil arabinoside Natural products OC1C(O)C(CO)OC1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-UHFFFAOYSA-N 0.000 description 1
- 229940045145 uridine Drugs 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 238000011179 visual inspection Methods 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
- 239000012138 yeast extract Substances 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
- C12N15/11—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
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of 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
- 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 [CRISPRs]
Landscapes
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201461931515P | 2014-01-24 | 2014-01-24 | |
| US61/931,515 | 2014-01-24 | ||
| PCT/US2015/012747 WO2015112896A2 (en) | 2014-01-24 | 2015-01-23 | Methods and compositions for sequences guiding cas9 targeting |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2936646A1 CA2936646A1 (en) | 2015-07-30 |
| CA2936646C true CA2936646C (en) | 2024-04-30 |
Family
ID=53682132
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2936646A Active CA2936646C (en) | 2014-01-24 | 2015-01-23 | Methods and compositions for sequences guiding cas9 targeting |
Country Status (4)
| Country | Link |
|---|---|
| EP (1) | EP3097212A4 (ja) |
| JP (4) | JP2017503514A (ja) |
| CA (1) | CA2936646C (ja) |
| WO (1) | WO2015112896A2 (ja) |
Families Citing this family (46)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3613852A3 (en) | 2011-07-22 | 2020-04-22 | President and Fellows of Harvard College | Evaluation and improvement of nuclease cleavage specificity |
| US9163284B2 (en) | 2013-08-09 | 2015-10-20 | President And Fellows Of Harvard College | Methods for identifying a target site of a Cas9 nuclease |
| CA3157565A1 (en) | 2013-08-22 | 2015-02-26 | E. I. Du Pont De Nemours And Company | Plant genome modification using guide rna/cas endonuclease systems and methods of use |
| US9359599B2 (en) | 2013-08-22 | 2016-06-07 | President And Fellows Of Harvard College | Engineered transcription activator-like effector (TALE) domains and uses thereof |
| US9388430B2 (en) | 2013-09-06 | 2016-07-12 | President And Fellows Of Harvard College | Cas9-recombinase fusion proteins and uses thereof |
| US9526784B2 (en) | 2013-09-06 | 2016-12-27 | President And Fellows Of Harvard College | Delivery system for functional nucleases |
| US9228207B2 (en) | 2013-09-06 | 2016-01-05 | President And Fellows Of Harvard College | Switchable gRNAs comprising aptamers |
| WO2015066119A1 (en) | 2013-10-30 | 2015-05-07 | North Carolina State University | Compositions and methods related to a type-ii crispr-cas system in lactobacillus buchneri |
| KR102523466B1 (ko) | 2013-11-07 | 2023-04-20 | 에디타스 메디신, 인코포레이티드 | 지배적인 gRNA를 이용하는 CRISPR-관련 방법 및 조성물 |
| US9068179B1 (en) | 2013-12-12 | 2015-06-30 | President And Fellows Of Harvard College | Methods for correcting presenilin point mutations |
| WO2015112896A2 (en) * | 2014-01-24 | 2015-07-30 | North Carolina State University | Methods and compositions for sequences guiding cas9 targeting |
| US11439712B2 (en) | 2014-04-08 | 2022-09-13 | North Carolina State University | Methods and compositions for RNA-directed repression of transcription using CRISPR-associated genes |
| CA2956224A1 (en) | 2014-07-30 | 2016-02-11 | President And Fellows Of Harvard College | Cas9 proteins including ligand-dependent inteins |
| WO2016033298A1 (en) | 2014-08-28 | 2016-03-03 | North Carolina State University | Novel cas9 proteins and guiding features for dna targeting and genome editing |
| BR112017017260A2 (pt) | 2015-03-27 | 2018-04-17 | Du Pont | construções de dna, vetor, célula, plantas, semente, método de expressão de rna e método de modificação de local alvo |
| CN107709562A (zh) * | 2015-05-15 | 2018-02-16 | 先锋国际良种公司 | 指导rna/cas内切核酸酶系统 |
| KR102451796B1 (ko) | 2015-05-29 | 2022-10-06 | 노쓰 캐롤라이나 스테이트 유니버시티 | 크리스퍼 핵산을 사용하여 박테리아, 고세균, 조류 및 효모를 스크리닝하는 방법 |
| WO2016205276A1 (en) | 2015-06-15 | 2016-12-22 | North Carolina State University | Methods and compositions for efficient delivery of nucleic acids and rna-based antimicrobials |
| US9580727B1 (en) | 2015-08-07 | 2017-02-28 | Caribou Biosciences, Inc. | Compositions and methods of engineered CRISPR-Cas9 systems using split-nexus Cas9-associated polynucleotides |
| WO2017058751A1 (en) | 2015-09-28 | 2017-04-06 | North Carolina State University | Methods and compositions for sequence specific antimicrobials |
| AU2016342380B2 (en) | 2015-10-23 | 2022-04-07 | President And Fellows Of Harvard College | Nucleobase editors and uses thereof |
| US11542466B2 (en) | 2015-12-22 | 2023-01-03 | North Carolina State University | Methods and compositions for delivery of CRISPR based antimicrobials |
| CN105594664B (zh) * | 2016-02-16 | 2018-10-02 | 湖南师范大学 | 一种基因敲除选育stat1a基因缺失型斑马鱼的方法 |
| GB2568182A (en) | 2016-08-03 | 2019-05-08 | Harvard College | Adenosine nucleobase editors and uses thereof |
| JP7201153B2 (ja) | 2016-08-09 | 2023-01-10 | プレジデント アンド フェローズ オブ ハーバード カレッジ | プログラム可能ｃａｓ９－リコンビナーゼ融合タンパク質およびその使用 |
| US11542509B2 (en) | 2016-08-24 | 2023-01-03 | President And Fellows Of Harvard College | Incorporation of unnatural amino acids into proteins using base editing |
| SG11201903089RA (en) | 2016-10-14 | 2019-05-30 | Harvard College | Aav delivery of nucleobase editors |
| WO2018119359A1 (en) | 2016-12-23 | 2018-06-28 | President And Fellows Of Harvard College | Editing of ccr5 receptor gene to protect against hiv infection |
| TW201839136A (zh) | 2017-02-06 | 2018-11-01 | 瑞士商諾華公司 | 治療血色素異常症之組合物及方法 |
| US11898179B2 (en) | 2017-03-09 | 2024-02-13 | President And Fellows Of Harvard College | Suppression of pain by gene editing |
| WO2018165629A1 (en) | 2017-03-10 | 2018-09-13 | President And Fellows Of Harvard College | Cytosine to guanine base editor |
| CA3057192A1 (en) | 2017-03-23 | 2018-09-27 | President And Fellows Of Harvard College | Nucleobase editors comprising nucleic acid programmable dna binding proteins |
| US11560566B2 (en) | 2017-05-12 | 2023-01-24 | President And Fellows Of Harvard College | Aptazyme-embedded guide RNAs for use with CRISPR-Cas9 in genome editing and transcriptional activation |
| WO2018217981A1 (en) | 2017-05-26 | 2018-11-29 | North Carolina State University | Altered guide rnas for modulating cas9 activity and methods of use |
| EP3652312A1 (en) | 2017-07-14 | 2020-05-20 | Editas Medicine, Inc. | Systems and methods for targeted integration and genome editing and detection thereof using integrated priming sites |
| CN111801345A (zh) | 2017-07-28 | 2020-10-20 | 哈佛大学的校长及成员们 | 使用噬菌体辅助连续进化(pace)的进化碱基编辑器的方法和组合物 |
| WO2019139645A2 (en) | 2017-08-30 | 2019-07-18 | President And Fellows Of Harvard College | High efficiency base editors comprising gam |
| CA3082251A1 (en) | 2017-10-16 | 2019-04-25 | The Broad Institute, Inc. | Uses of adenosine base editors |
| WO2019165168A1 (en) | 2018-02-23 | 2019-08-29 | Pioneer Hi-Bred International, Inc. | Novel cas9 orthologs |
| US10711267B2 (en) | 2018-10-01 | 2020-07-14 | North Carolina State University | Recombinant type I CRISPR-Cas system |
| CA3117228A1 (en) | 2018-12-14 | 2020-06-18 | Pioneer Hi-Bred International, Inc. | Novel crispr-cas systems for genome editing |
| SG11202109679VA (en) | 2019-03-19 | 2021-10-28 | Broad Inst Inc | Methods and compositions for editing nucleotide sequences |
| CN110117622B (zh) * | 2019-05-24 | 2021-05-11 | 青岛农业大学 | 一种CRISPR/Cas基因编辑系统及其制备方法和应用 |
| WO2020263773A1 (en) * | 2019-06-24 | 2020-12-30 | North Carolina State University | Methods and compositions for modifying bacterial genomes and for killing bacteria |
| WO2021216772A1 (en) * | 2020-04-21 | 2021-10-28 | Mammoth Biosciences, Inc. | Casy programmable nucleases and rna component systems |
| MX2022014008A (es) | 2020-05-08 | 2023-02-09 | Broad Inst Inc | Métodos y composiciones para la edición simultánea de ambas cadenas de una secuencia de nucleótidos de doble cadena objetivo. |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DK3401400T3 (da) * | 2012-05-25 | 2019-06-03 | Univ California | Fremgangsmåder og sammensætninger til rna-styret mål-dna-modifikation og til rna-styret transskriptionsmodulering |
| WO2015112896A2 (en) * | 2014-01-24 | 2015-07-30 | North Carolina State University | Methods and compositions for sequences guiding cas9 targeting |
-
2015
- 2015-01-23 WO PCT/US2015/012747 patent/WO2015112896A2/en active Application Filing
- 2015-01-23 CA CA2936646A patent/CA2936646C/en active Active
- 2015-01-23 JP JP2016547162A patent/JP2017503514A/ja active Pending
- 2015-01-23 EP EP15740979.8A patent/EP3097212A4/en active Pending
-
2020
- 2020-02-13 JP JP2020022160A patent/JP2020103295A/ja active Pending
-
2022
- 2022-04-28 JP JP2022074741A patent/JP7429057B2/ja active Active
-
2024
- 2024-01-16 JP JP2024004564A patent/JP2024041934A/ja active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| CA2936646A1 (en) | 2015-07-30 |
| EP3097212A2 (en) | 2016-11-30 |
| EP3097212A4 (en) | 2017-10-04 |
| JP2020103295A (ja) | 2020-07-09 |
| JP2024041934A (ja) | 2024-03-27 |
| WO2015112896A9 (en) | 2015-11-26 |
| JP2017503514A (ja) | 2017-02-02 |
| JP7429057B2 (ja) | 2024-02-07 |
| WO2015112896A3 (en) | 2015-10-29 |
| WO2015112896A2 (en) | 2015-07-30 |
| JP2022106883A (ja) | 2022-07-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20210017507A1 (en) | Methods and compositions for sequences guiding cas9 targeting | |
| CA2936646C (en) | Methods and compositions for sequences guiding cas9 targeting | |
| US11753651B2 (en) | Cas9 proteins and guiding features for DNA targeting and genome editing | |
| US11499169B2 (en) | Compositions and methods related to a type-II CRISPR-Cas system in Lactobacillus buchneri | |
| US20200263186A1 (en) | Altered guide rnas for modulating cas9 activity and methods of use | |
| AU2022204596A1 (en) | Methods for screening bacteria, archaea, algae, and yeast using crispr nucleic acids | |
| KR20220110739A (ko) | 유형 v crispr-cas 염기 편집제 및 그의 사용 방법 | |
| WO2021081367A1 (en) | Compositions and methods for rna-templated editing in plants | |
| CN116745415A (zh) | 工程化crispr-cas蛋白质及其使用方法 | |
| CN115380111A (zh) | 用于碱基多样化的组合物、系统和方法 | |
| CA3187135A1 (en) | Compositions, systems, and methods for base diversification | |
| WO2024017189A1 (en) | Tnpb-based genome editor | |
| US20230051466A1 (en) | crRNA:tracrRNA-BASED BINARY LOGIC GATE DESIGN AS A TOOL FOR SYNTHETIC BIOLOGY | |
| WO2023039565A2 (en) | Methods for improving phage resistance in clostridium species | |
| WO2023164722A1 (en) | Engineered crispr-cas effector proteins and methods of use thereof | |
| WO2024182746A1 (en) | Engineered proteins and methods of use thereof | |
| CA3161392A1 (en) | Codon-optimized cas9 endonuclease encoding polynucleotide | |
| CN118726313A (en) | CAS9 mutant gene of Streptococcus pyogenes and polypeptide encoded thereby |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request |
Effective date: 20191224 |
|
| EEER | Examination request |
Effective date: 20191224 |
|
| EEER | Examination request |
Effective date: 20191224 |
|
| EEER | Examination request |
Effective date: 20191224 |
|
| EEER | Examination request |
Effective date: 20191224 |
|
| EEER | Examination request |
Effective date: 20191224 |
|
| EEER | Examination request |
Effective date: 20191224 |
|
| EEER | Examination request |
Effective date: 20191224 |
|
| EEER | Examination request |
Effective date: 20191224 |