CN114269941A - 基于脱靶评估评价基因编辑治疗的方法 - Google Patents
基于脱靶评估评价基因编辑治疗的方法 Download PDFInfo
- Publication number
- CN114269941A CN114269941A CN202080058282.4A CN202080058282A CN114269941A CN 114269941 A CN114269941 A CN 114269941A CN 202080058282 A CN202080058282 A CN 202080058282A CN 114269941 A CN114269941 A CN 114269941A
- Authority
- CN
- China
- Prior art keywords
- target
- genetically modified
- site
- gene editing
- target marker
- 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.)
- Pending
Links
- 238000000034 method Methods 0.000 title claims abstract description 273
- 238000010362 genome editing Methods 0.000 title claims abstract description 174
- 238000002560 therapeutic procedure Methods 0.000 title claims abstract description 27
- 238000012911 target assessment Methods 0.000 title description 2
- 238000002659 cell therapy Methods 0.000 claims abstract description 56
- 238000004458 analytical method Methods 0.000 claims abstract description 21
- 239000003550 marker Substances 0.000 claims description 161
- 102100022976 B-cell lymphoma/leukemia 11A Human genes 0.000 claims description 80
- 101000903703 Homo sapiens B-cell lymphoma/leukemia 11A Proteins 0.000 claims description 80
- 108091033409 CRISPR Proteins 0.000 claims description 55
- 150000007523 nucleic acids Chemical class 0.000 claims description 38
- 108020004707 nucleic acids Proteins 0.000 claims description 27
- 102000039446 nucleic acids Human genes 0.000 claims description 27
- 238000010354 CRISPR gene editing Methods 0.000 claims description 25
- 230000008859 change Effects 0.000 claims description 24
- 238000003776 cleavage reaction Methods 0.000 claims description 23
- 230000007017 scission Effects 0.000 claims description 23
- 238000010453 CRISPR/Cas method Methods 0.000 claims description 21
- 238000000099 in vitro assay Methods 0.000 claims description 21
- 238000000338 in vitro Methods 0.000 claims description 20
- 238000012163 sequencing technique Methods 0.000 claims description 17
- 238000010998 test method Methods 0.000 claims description 14
- 238000009738 saturating Methods 0.000 claims description 10
- 238000001712 DNA sequencing Methods 0.000 claims description 8
- 230000008030 elimination Effects 0.000 claims description 7
- 238000003379 elimination reaction Methods 0.000 claims description 7
- 238000012544 monitoring process Methods 0.000 abstract description 8
- 238000003766 bioinformatics method Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 199
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 62
- 239000003623 enhancer Substances 0.000 description 43
- 101100493741 Homo sapiens BCL11A gene Proteins 0.000 description 32
- 125000003729 nucleotide group Chemical group 0.000 description 25
- 239000002773 nucleotide Substances 0.000 description 24
- 210000000130 stem cell Anatomy 0.000 description 24
- 108090000623 proteins and genes Proteins 0.000 description 22
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 20
- 108010054147 Hemoglobins Proteins 0.000 description 18
- 102000001554 Hemoglobins Human genes 0.000 description 18
- 201000010099 disease Diseases 0.000 description 18
- 230000009437 off-target effect Effects 0.000 description 17
- 108020004414 DNA Proteins 0.000 description 16
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 description 16
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 description 16
- 108091028043 Nucleic acid sequence Proteins 0.000 description 15
- 230000014509 gene expression Effects 0.000 description 15
- 210000000349 chromosome Anatomy 0.000 description 13
- 210000003743 erythrocyte Anatomy 0.000 description 13
- 241000282414 Homo sapiens Species 0.000 description 12
- 230000000694 effects Effects 0.000 description 12
- 230000008569 process Effects 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 12
- 238000001514 detection method Methods 0.000 description 11
- 230000004069 differentiation Effects 0.000 description 11
- 230000000925 erythroid effect Effects 0.000 description 11
- 238000012350 deep sequencing Methods 0.000 description 10
- 230000006870 function Effects 0.000 description 10
- 238000012239 gene modification Methods 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 230000005782 double-strand break Effects 0.000 description 8
- 230000002829 reductive effect Effects 0.000 description 8
- 230000003394 haemopoietic effect Effects 0.000 description 7
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 6
- 108010044495 Fetal Hemoglobin Proteins 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 230000008685 targeting Effects 0.000 description 6
- 108091005886 Hemoglobin subunit gamma Proteins 0.000 description 5
- 238000010459 TALEN Methods 0.000 description 5
- 208000002903 Thalassemia Diseases 0.000 description 5
- 108010043645 Transcription Activator-Like Effector Nucleases Proteins 0.000 description 5
- 208000005980 beta thalassemia Diseases 0.000 description 5
- 210000001185 bone marrow Anatomy 0.000 description 5
- 230000001413 cellular effect Effects 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 230000000295 complement effect Effects 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 230000004048 modification Effects 0.000 description 5
- 238000012986 modification Methods 0.000 description 5
- 230000035772 mutation Effects 0.000 description 5
- 210000005259 peripheral blood Anatomy 0.000 description 5
- 239000011886 peripheral blood Substances 0.000 description 5
- 208000024891 symptom Diseases 0.000 description 5
- 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 4
- 102100038617 Hemoglobin subunit gamma-2 Human genes 0.000 description 4
- 108010090804 Streptavidin Proteins 0.000 description 4
- 108010017070 Zinc Finger Nucleases Proteins 0.000 description 4
- 230000003321 amplification Effects 0.000 description 4
- 239000002246 antineoplastic agent Substances 0.000 description 4
- 230000027455 binding Effects 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 238000002512 chemotherapy Methods 0.000 description 4
- 238000005520 cutting process Methods 0.000 description 4
- 229940127089 cytotoxic agent Drugs 0.000 description 4
- 230000007423 decrease Effects 0.000 description 4
- 210000003013 erythroid precursor cell Anatomy 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 230000002068 genetic effect Effects 0.000 description 4
- 230000005017 genetic modification Effects 0.000 description 4
- 235000013617 genetically modified food Nutrition 0.000 description 4
- 238000003199 nucleic acid amplification method Methods 0.000 description 4
- 230000035755 proliferation Effects 0.000 description 4
- 210000001995 reticulocyte Anatomy 0.000 description 4
- 238000010356 CRISPR-Cas9 genome editing Methods 0.000 description 3
- 108091005904 Hemoglobin subunit beta Proteins 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- 108091028113 Trans-activating crRNA Proteins 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 230000032823 cell division Effects 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 238000001415 gene therapy Methods 0.000 description 3
- 210000003917 human chromosome Anatomy 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 238000003780 insertion Methods 0.000 description 3
- 230000037431 insertion Effects 0.000 description 3
- 238000002372 labelling Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 230000005945 translocation Effects 0.000 description 3
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 2
- COVZYZSDYWQREU-UHFFFAOYSA-N Busulfan Chemical compound CS(=O)(=O)OCCCCOS(C)(=O)=O COVZYZSDYWQREU-UHFFFAOYSA-N 0.000 description 2
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 102000004269 Granulocyte Colony-Stimulating Factor Human genes 0.000 description 2
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 description 2
- 101150086355 HBG gene Proteins 0.000 description 2
- 101001105486 Homo sapiens Proteasome subunit alpha type-7 Proteins 0.000 description 2
- 101710163270 Nuclease Proteins 0.000 description 2
- 102100021201 Proteasome subunit alpha type-7 Human genes 0.000 description 2
- 241000193996 Streptococcus pyogenes Species 0.000 description 2
- 101000910035 Streptococcus pyogenes serotype M1 CRISPR-associated endonuclease Cas9/Csn1 Proteins 0.000 description 2
- 108091023040 Transcription factor Proteins 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 210000000601 blood cell Anatomy 0.000 description 2
- 229960002092 busulfan Drugs 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 230000011748 cell maturation Effects 0.000 description 2
- 238000007385 chemical modification Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 229960004397 cyclophosphamide Drugs 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 230000001605 fetal effect Effects 0.000 description 2
- 229960000390 fludarabine Drugs 0.000 description 2
- GIUYCYHIANZCFB-FJFJXFQQSA-N fludarabine phosphate Chemical compound C1=NC=2C(N)=NC(F)=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@@H]1O GIUYCYHIANZCFB-FJFJXFQQSA-N 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 108010038853 gamma-Globins Proteins 0.000 description 2
- 210000005260 human cell Anatomy 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000002625 monoclonal antibody therapy Methods 0.000 description 2
- 230000001400 myeloablative effect Effects 0.000 description 2
- 210000003924 normoblast Anatomy 0.000 description 2
- 210000004940 nucleus Anatomy 0.000 description 2
- 210000003463 organelle Anatomy 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 108091033319 polynucleotide Proteins 0.000 description 2
- 102000040430 polynucleotide Human genes 0.000 description 2
- 239000002157 polynucleotide Substances 0.000 description 2
- 230000002062 proliferating effect Effects 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 208000007056 sickle cell anemia Diseases 0.000 description 2
- 239000000344 soap Substances 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 125000000446 sulfanediyl group Chemical group *S* 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- 230000009885 systemic effect Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- PZNPLUBHRSSFHT-RRHRGVEJSA-N 1-hexadecanoyl-2-octadecanoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)O[C@@H](COP([O-])(=O)OCC[N+](C)(C)C)COC(=O)CCCCCCCCCCCCCCC PZNPLUBHRSSFHT-RRHRGVEJSA-N 0.000 description 1
- 102100035248 Alpha-(1,3)-fucosyltransferase 4 Human genes 0.000 description 1
- 108091093088 Amplicon Proteins 0.000 description 1
- 241000203069 Archaea Species 0.000 description 1
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 description 1
- 108050005493 CD3 protein, epsilon/gamma/delta subunit Proteins 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 102100035716 Glycophorin-A Human genes 0.000 description 1
- 108060003393 Granulin Proteins 0.000 description 1
- 108020005004 Guide RNA Proteins 0.000 description 1
- 102100021519 Hemoglobin subunit beta Human genes 0.000 description 1
- 102100039894 Hemoglobin subunit delta Human genes 0.000 description 1
- 108091005903 Hemoglobin subunit delta Proteins 0.000 description 1
- 108091005879 Hemoglobin subunit epsilon Proteins 0.000 description 1
- 208000035185 Hemolytic Congenital Anemia Diseases 0.000 description 1
- 206010060893 Hereditary haemolytic anaemia Diseases 0.000 description 1
- 102000006947 Histones Human genes 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 101001022185 Homo sapiens Alpha-(1,3)-fucosyltransferase 4 Proteins 0.000 description 1
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 1
- 101001074244 Homo sapiens Glycophorin-A Proteins 0.000 description 1
- 101000899111 Homo sapiens Hemoglobin subunit beta Proteins 0.000 description 1
- 101001046686 Homo sapiens Integrin alpha-M Proteins 0.000 description 1
- 101000998120 Homo sapiens Interleukin-3 receptor subunit alpha Proteins 0.000 description 1
- 101000878605 Homo sapiens Low affinity immunoglobulin epsilon Fc receptor Proteins 0.000 description 1
- 101000917858 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-A Proteins 0.000 description 1
- 101000917839 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-B Proteins 0.000 description 1
- 101000946889 Homo sapiens Monocyte differentiation antigen CD14 Proteins 0.000 description 1
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 description 1
- 101000934346 Homo sapiens T-cell surface antigen CD2 Proteins 0.000 description 1
- 102100022338 Integrin alpha-M Human genes 0.000 description 1
- 102100033493 Interleukin-3 receptor subunit alpha Human genes 0.000 description 1
- 102100038007 Low affinity immunoglobulin epsilon Fc receptor Human genes 0.000 description 1
- 102100029185 Low affinity immunoglobulin gamma Fc region receptor III-B Human genes 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 102100035877 Monocyte differentiation antigen CD14 Human genes 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 description 1
- 101100396537 Solanum lycopersicum eIF4E1 gene Proteins 0.000 description 1
- 102100025237 T-cell surface antigen CD2 Human genes 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 241000283907 Tragelaphus oryx Species 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 201000006288 alpha thalassemia Diseases 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 239000007640 basal medium Substances 0.000 description 1
- 102000023732 binding proteins Human genes 0.000 description 1
- 108091008324 binding proteins Proteins 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 201000001516 congenital hemolytic anemia Diseases 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 230000008029 eradication Effects 0.000 description 1
- 210000000267 erythroid cell Anatomy 0.000 description 1
- 208000024987 familial hemolytic anemia Diseases 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 108091008053 gene clusters Proteins 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 210000002288 golgi apparatus Anatomy 0.000 description 1
- 210000000777 hematopoietic system Anatomy 0.000 description 1
- 238000012165 high-throughput sequencing Methods 0.000 description 1
- 210000001624 hip Anatomy 0.000 description 1
- 210000001981 hip bone Anatomy 0.000 description 1
- 230000007124 immune defense Effects 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 210000003738 lymphoid progenitor cell Anatomy 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 230000001483 mobilizing effect Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 210000000633 nuclear envelope Anatomy 0.000 description 1
- 229940046166 oligodeoxynucleotide Drugs 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 230000003169 placental effect Effects 0.000 description 1
- 230000004983 pleiotropic effect Effects 0.000 description 1
- 210000002729 polyribosome Anatomy 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 230000001902 propagating effect Effects 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 210000000614 rib Anatomy 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 210000001562 sternum Anatomy 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/90—Stable introduction of foreign DNA into chromosome
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
-
- 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]
-
- 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
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6869—Methods for sequencing
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Developmental Biology & Embryology (AREA)
- Pathology (AREA)
- Hematology (AREA)
- Plant Pathology (AREA)
- Virology (AREA)
- Mycology (AREA)
- Urology & Nephrology (AREA)
- Gastroenterology & Hepatology (AREA)
- Food Science & Technology (AREA)
- Toxicology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Chemical Kinetics & Catalysis (AREA)
Abstract
一种确定细胞疗法对个体的适合性的方法。利用生物信息学方法和无偏向的潜在脱靶位点实验分析方法,在向个体施用细胞疗法前对源自个体的基因修饰细胞的潜在脱靶位点进行预测,并在将源自个体的基因修饰细胞施用至个体后对潜在脱靶位点进行监测,确定所述潜在脱靶位点否发生基因编辑,并基于基因编辑事件的发生评估所述治疗的适合性。
Description
PCT国内申请,说明书已公开。
Claims (35)
- PCT国内申请,权利要求书已公开。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CNPCT/CN2019/104303 | 2019-09-04 | ||
CN2019104303 | 2019-09-04 | ||
PCT/CN2020/113562 WO2021043278A1 (zh) | 2019-09-04 | 2020-09-04 | 基于脱靶评估评价基因编辑治疗的方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN114269941A true CN114269941A (zh) | 2022-04-01 |
Family
ID=74852757
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202080058282.4A Pending CN114269941A (zh) | 2019-09-04 | 2020-09-04 | 基于脱靶评估评价基因编辑治疗的方法 |
Country Status (8)
Country | Link |
---|---|
US (1) | US20220340969A1 (zh) |
EP (1) | EP4026910A1 (zh) |
JP (1) | JP2022547105A (zh) |
KR (1) | KR20220053671A (zh) |
CN (1) | CN114269941A (zh) |
AU (1) | AU2020342207A1 (zh) |
CA (1) | CA3150230A1 (zh) |
WO (1) | WO2021043278A1 (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20220307020A1 (en) | 2019-04-15 | 2022-09-29 | Edigene Inc. | Methods and compositions for editing rnas |
EP3997229A4 (en) | 2019-07-12 | 2024-07-03 | Univ Beijing | TARGETED RNA EDITING BY HARNESSING ENDOGENOUS ADAR USING MODIFIED RNA |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109735574A (zh) * | 2017-10-27 | 2019-05-10 | 博雅辑因(北京)生物科技有限公司 | 一种提高胎儿血红蛋白表达的方法 |
CN109789185A (zh) * | 2016-07-28 | 2019-05-21 | 基础科学研究院 | 用于治疗眼部疾病的含cas9蛋白和向导rna的药物组合物 |
Family Cites Families (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
HUE038850T2 (hu) | 2012-05-25 | 2018-11-28 | Univ California | Eljárások és kompozíciók cél-DNS RNS-irányított módosításához és transzkripció RNS-irányított modulálásához |
AU2013293270B2 (en) | 2012-07-25 | 2018-08-16 | Massachusetts Institute Of Technology | Inducible DNA binding proteins and genome perturbation tools and applications thereof |
CN110066775B (zh) | 2012-10-23 | 2024-03-19 | 基因工具股份有限公司 | 用于切割靶dna的组合物及其用途 |
US8697359B1 (en) | 2012-12-12 | 2014-04-15 | The Broad Institute, Inc. | CRISPR-Cas systems and methods for altering expression of gene products |
CN108351350B (zh) * | 2015-08-25 | 2022-02-18 | 杜克大学 | 使用rna指导型内切核酸酶改善基因组工程特异性的组合物和方法 |
EP3450570B1 (en) * | 2016-04-28 | 2021-03-10 | Industry-Academic Cooperation Foundation, Yonsei University | Method for evaluating, in vivo, activity of rna-guided nuclease in high-throughput manner |
MX2018013445A (es) * | 2016-05-06 | 2019-09-09 | Juno Therapeutics Inc | Celulas diseñadas geneticamente y metodos para obtener las mismas. |
KR102558931B1 (ko) * | 2017-06-23 | 2023-07-21 | 인스크립타 인코포레이티드 | 핵산 가이드 뉴클레아제 |
WO2019052577A1 (zh) * | 2017-09-18 | 2019-03-21 | 博雅辑因(北京)生物科技有限公司 | 一种基因编辑t细胞及其用途 |
EP3697907A1 (en) * | 2017-10-17 | 2020-08-26 | CRISPR Therapeutics AG | Compositions and methods for gene editing for hemophilia a |
CN107967411B (zh) * | 2017-11-21 | 2021-09-10 | 南方科技大学 | 一种脱靶位点的检测方法、装置及终端设备 |
CN109868283B (zh) * | 2019-02-21 | 2021-07-20 | 浙江农林大学 | 一种评估CRISPR/Cas9基因编辑效率或脱靶频率的方法 |
-
2020
- 2020-09-04 CA CA3150230A patent/CA3150230A1/en active Pending
- 2020-09-04 WO PCT/CN2020/113562 patent/WO2021043278A1/zh unknown
- 2020-09-04 CN CN202080058282.4A patent/CN114269941A/zh active Pending
- 2020-09-04 KR KR1020227011062A patent/KR20220053671A/ko not_active Application Discontinuation
- 2020-09-04 EP EP20861118.6A patent/EP4026910A1/en not_active Withdrawn
- 2020-09-04 JP JP2022514832A patent/JP2022547105A/ja active Pending
- 2020-09-04 AU AU2020342207A patent/AU2020342207A1/en active Pending
- 2020-09-04 US US17/640,759 patent/US20220340969A1/en active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109789185A (zh) * | 2016-07-28 | 2019-05-21 | 基础科学研究院 | 用于治疗眼部疾病的含cas9蛋白和向导rna的药物组合物 |
CN109735574A (zh) * | 2017-10-27 | 2019-05-10 | 博雅辑因(北京)生物科技有限公司 | 一种提高胎儿血红蛋白表达的方法 |
Non-Patent Citations (1)
Title |
---|
王影等: "CRISPR/Cas9基因组定点编辑中脱靶现象的研究进展", 植物学报, vol. 53, no. 4, pages 528 - 541 * |
Also Published As
Publication number | Publication date |
---|---|
US20220340969A1 (en) | 2022-10-27 |
KR20220053671A (ko) | 2022-04-29 |
WO2021043278A1 (zh) | 2021-03-11 |
EP4026910A1 (en) | 2022-07-13 |
JP2022547105A (ja) | 2022-11-10 |
CA3150230A1 (en) | 2021-03-11 |
AU2020342207A1 (en) | 2022-04-21 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN109735497B (zh) | 一种提高胎儿血红蛋白表达水平的方法 | |
Schumann et al. | Functional CRISPR dissection of gene networks controlling human regulatory T cell identity | |
Vionnie et al. | Epigenetic memory underlies cell-autonomous heterogeneous behavior of hematopoietic stem cells | |
EP3964579A1 (en) | Method for predicting effectiveness of treatment of hemoglobinopathy | |
CN114269941A (zh) | 基于脱靶评估评价基因编辑治疗的方法 | |
Shi et al. | Clonal expansion and myeloid leukemia progression modeled by multiplex gene editing of murine hematopoietic progenitor cells | |
Zhang et al. | Ex vivo expansion potential of murine hematopoietic stem cells is a rare property only partially predicted by phenotype | |
Martin et al. | Dynamics of chromatin accessibility during hematopoietic stem cell differentiation into progressively lineage-committed progeny | |
Yang et al. | Tumor‐Associated Monocytes Reprogram CD8+ T Cells into Central Memory‐Like Cells with Potent Antitumor Effects | |
CN112447264A (zh) | 基于脱靶评估评价基因编辑治疗的方法 | |
CN111939271A (zh) | 一种血红蛋白病治疗有效性预测方法 | |
Shannon | The Molecular Biology of Sickle Cell Anaemia | |
CA3237748A1 (en) | Biomarkers of megakaryocyte-derived extracellular vesicles | |
CN114222816A (zh) | 一种高效修复环状铁粒幼细胞性贫血基因突变的方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
TA01 | Transfer of patent application right |
Effective date of registration: 20220712 Address after: 510000 building 10, No.6 Nanjiang 2nd Road, Zhujiang street, Nansha District, Guangzhou City, Guangdong Province Applicant after: Guangzhou Ji Yin Medical Technology Co.,Ltd. Address before: 102206 2nd floor, building 2, No.22, kekeyuan Road, Changping District, Beijing Applicant before: EDIGENE Inc. |
|
TA01 | Transfer of patent application right | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20220401 |
|
WD01 | Invention patent application deemed withdrawn after publication |