基因编辑技术在人类生殖细胞中的应用研究
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摘要
基因编辑技术极大地提高了对基因组靶位点精确修改的效率。人类早期胚胎发育的特异性和目前报道的上万种遗传突变的修复必要性使得基因编辑技术在人生殖细胞中的应用成为必然。对人生殖细胞基因编辑是一个新的领域,涉及到伦理的考虑。因此,这既是个科学问题,同时也是个社会问题。现综述基因编辑技术在人类生殖细胞应用的伦理之争和研究进展。
Genome editing greatly promotes the efficiency of precise modification of target sites. It is necessary to edit the germline cell because of the specificity of human embryo development and the necessity for repairing thousands of genetic diseases. It is a new field to edit the germline cell, and it is not only a scientific problem, but also an ethical issue. This article summarizes the ethical discussion of germline cell editing and its research progress.
Genome editing greatly promotes the efficiency of precise modification of target sites. It is necessary to edit the germline cell because of the specificity of human embryo development and the necessity for repairing thousands of genetic diseases. It is a new field to edit the germline cell, and it is not only a scientific problem, but also an ethical issue. This article summarizes the ethical discussion of germline cell editing and its research progress.
引文
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[2]Cong L,Ran FA,Cox D,et al.Multiplex genome engineering using CRISPR/Cas systems.Science,2013,339:819-3
[3]Komor AC,Badran AH,Liu DR.CRISPR-based technologies for the manipulation of eukaryotic genomes.Cell,2017,168:20-36
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[5]Petropoulos S,Edsgard D,Reinius B,et al.Single-cell RNA-Seq reveals lineage and X chromosome dynamics in human preimplantation embryos.Cell,2016,165:1012-26
[6]Cyranoski D,Reardon S.Embryo editing sparks epic debate.Nature,2015,520:593-4
[7]Ma H,Marti-Gutierrez N,Park SW,et al.Correction of a pathogenic gene mutation in human embryos.Nature,2017,548:413-9
[8]Pei D,Beier DW,Levy-Lahad E,et al.Human embryo editing:opportunities and importance of transnational cooperation.Cell Stem Cell,2017,21:423-6
[9]Vog G.Embryo edit makes human‘knockout’.Science,2017,357:1225
[10]Rossant J.Gene editing in human development:ethical concerns and practical applications.Development,2018,145,pii:dev150888
[11]Callaway E.Gene-editing research in human embryos gains momentum.Nature,2016,532:289-90
[12]Liang P,Xu Y,Zhang X,et al.CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes.Protein Cell,2015,6:363-72
[13]Ishii T.Germ line genome editing in clinics:the approaches,objectives and global society.Brief Funct Genomics,2017,16:46-56
[14]Li G,Liu Y,Zeng Y,et al.Highly efficient and precise base editing in discarded human tripronuclear embryos.Protein Cell,2017,8:776-9
[15]Kang XJ,He WY,Huang YL,et al.Introducing precise genetic modifications into human 3PN embryos by CRIS-PR/Cas-mediated genome editing.J Assist Reprod Genet,2016,33:581-8
[16]Fogarty NM,McCarthy A,Snijders KE,et al.Genome editing reveals a role for OCT4 in human embryogenesis.Nature,2017,550:67-73
[17]Tang L,Zeng Y,Du H,et al.CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein.Mol Genet Genomics,2017,292:525-33
[18]Ruzo A,Brivanlou AH.At last:gene editing in human embryos to understand human development.Cell Stem Cell,2017,21:564-5
[19]Cyranoski D.Ethics of embryo editing divides scientists.Nature,2015,519:272
[20]Baltimore D,Berg P,Botchan M,et al.Biotechnology.Aprudent path forward for genomic engineering and germline gene modification.Science,2015,348:36-8
[21]Martikainen M,Pedersen O.Germline edits:heat does not help debate.Nature,2015,520:623
[22]Savulescu J,Gyngell C,Douglas T.Germline edits:trust ethics review process.Nature,2015,520:623
[23]Porteus MH,Dann CT.Genome editing of the germline:broadening the discussion.Mol Ther,2015,23:980-2
[24]Friedmann T,Jonlin EC,King NMP,et al.ASGCT and JSGT joint position statement on human genomic editing.Mol Ther,2015,23:1282
[25]Lanphier E,Urnov F,Haecker SE,et al.Don’t edit the human germ line.Nature,2015,519:410-1
[26]Miller HI.Germline gene therapy:we’re ready.Science,2015,348:1325
[27]Lunshof JE.Human germ line editing-roles and responsibilities.Protein Cell,2016,7:7-10
[28]Addison C,Taylor-Alexander S.Gene editing and germline intervention:the need for novel responses to novel technologies.Mol Ther,2015,23:1678-80
[29]Thompson C.Human embryos:collect reliable data on embryo selection.Nature,2017,551:33
[30]Regalado A.Engineering the perfect baby.Technol Rev,2015,118:26-33
[31]Cartier-Lacave N,Ali R,Yla-Herttuala S,et al.Debate on germline gene editing.Hum Gene Ther Methods,2016,27:135-42
[32]Ishii T.Reproductive medicine involving genome editing:clinical uncertainties and embryological needs.Reprod Biomed Online,2017,34:27-31
[33]Savulescu J,Pugh J,Douglas T,et al.The moral imperative to continue gene editing research on human embryos.Protein Cell,2015,6:476-9
[34]Ball P.Kathy Niakan:at the forefront of gene editing in embryos.Lancet,2016,387:935-5
[35]Kaiser J.A yellow light for embryo editing.Science,2017,355:675
[36]范月蕾,王慧媛,于建荣.基因编辑的伦理争议.科技中国,2018:98-104
[37]Ormond KE,Mortlock DP,Scholes DT,et al.Human germline genome editing.Am J Hum Genet,2017,101:167-76
[38]Plaza Reyes A,Lanner F.Towards a CRISPR view of early human development:applications,limitations and ethical concerns of genome editing in human embryos.Development,2017,144:3-7
[39]Human embryo research policy update.Nat Biotechnol,2018,36:477
[40]Austin CP,Dawkins HJS.Medical research:next decade’s goals for rare diseases.Nature,2017,548:158
[41]Hsu PD,Lander ES,Zhang F.Development and applications of CRISPR-Cas9 for genome engineering.Cell,2014,157:1262-78
[42]Gross M.Bacterial scissors to edit human embryos?Curr Biol,2015,25:R439-42
[43]Ledford H.CRISPR fixes disease gene in viable human embryos.Nature,2017,548:13-4
[44]Egli D,Zuccaro MV,Kosicki M,et al.Inter-homologue repair in fertilized human eggs?Nature,2018,560:E5-E7
[45]Schenkwein D,Yla-Herttuala S.Gene editing of human embryos with CRISPR/Cas9:Great promise coupled with important caveats.Mol Ther,2018,26:659-60
[46]Kosicki M,Tomberg K,Bradley A.Repair of double-strand breaks induced by CRISPR-Cas9 leads to large deletions and complex rearrangements.Nat Biotechnol,2018,36:765-71
[47]Haapaniemi E,Botla S,Persson J,et al.CRISPR-Cas9genome editing induces a p53-mediated DNA damage response.Nat Med,2018,24:927-30
[48]Ihry RJ,Worringer KA,Salick MR,et al.p53 inhibits CRISPR-Cas9 engineering in human pluripotent stem cells.Nat Med,2018,24:939-46
[49]Eid A,Alshareef S,Mahfouz MM.CRISPR base editors:genome editing without double-stranded breaks.Biochem J,2018,475:1955-64
[50]Hess GT,Tycko J,Yao D,et al.Methods and applications of CRISPR-mediated base editing in eukaryotic genomes.Mol Cell,2017,68:26-43
[51]Komor AC,Kim YB,Packer MS,et al.Programmable editing of a target base in genomic DNA without doublestranded DNA cleavage.Nature,2016,533:420-4
[52]Kim D,Lim K,Kim ST,et al.Genome-wide target specificities of CRISPR RNA-guided programmable deaminases.Nat Biotechnol,2017,35:475-80
[53]Koblan LW,Doman JL,Wilson C,et al.Improving cytidine and adenine base editors by expression optimization and ancestral reconstruction.Nat Biotechnol,2018,36:843-6
[54]Li X,Wang Y,Liu Y,et al.Base editing with a Cpf1-cytidine deaminase fusion.Nat Biotechnol,2018,36:324-7
[55]Wang X,Li J,Wang Y,et al.Efficient base editing in methylated regions with a human APOBEC3A-Cas9fusion.Nat Biotechnol,2018[Epub ahead of print]
[56]Kim YB,Komor AC,Levy JM,et al.Increasing the genome-targeting scope and precision of base editing with engineered Cas9-cytidine deaminase fusions.Nat Biotechnol,2017,35:371-6
[57]Jiang W,Feng S,Huang S,et al.BE-PLUS:a new base editing tool with broadened editing window and enhanced fidelity.Cell Res,2018[Epub ahead of print]
[58]Gaudelli NM,Komor AC,Rees HA,et al.Programmable base editing of A*T to G*C in genomic DNA without DNA cleavage.Nature,2017,551:464-71
[59]Liu Z,Chen M,Chen S,et al.Highly efficient RNA-guided base editing in rabbit.Nat Commun,2018,9:2717
[60]Li C,Zong Y,Wang Y,et al.Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion.Genome Biol,2018,19:59
[61]Liu Z,Lu Z,Yang G,et al.Efficient generation of mouse models of human diseases via ABE-and BE-mediated base editing.Nat Commun,2018,9:2338
[62]Zhou C,Zhang M,Wei Y,et al.Highly efficient base editing in human tripronuclear zygotes.Protein Cell,2017,8:772-5
[63]Liang PP,Ding CH,Sun HW,et al.Correction ofβ-thalassemia mutant by base editor in human embryos.Protein Cell,2017,8:811-22
[64]Zeng Y,Li J,Li G,et al.Correction of the Marfan syndrome pathogenic FBN1 mutation by base editing in human cells and heterozygous embryos.Mol Therapy,2018.pii:S1525-0016(18)30378-2
[65]Aird EJ,Lovendahl KN,St.Martin A,et al.Increasing Cas9-mediated homology-directed repair efficiency through covalent tethering of DNA repair template.Commun Biol,2018,1:54
[2]Cong L,Ran FA,Cox D,et al.Multiplex genome engineering using CRISPR/Cas systems.Science,2013,339:819-3
[3]Komor AC,Badran AH,Liu DR.CRISPR-based technologies for the manipulation of eukaryotic genomes.Cell,2017,168:20-36
[4]Cooper ML,Choi J,Staser K,et al.An“off-the-shelf”fratricide-resistant CAR-T for the treatment of T cell hematologic malignancies.Leukemia,2018,32:1970-83
[5]Petropoulos S,Edsgard D,Reinius B,et al.Single-cell RNA-Seq reveals lineage and X chromosome dynamics in human preimplantation embryos.Cell,2016,165:1012-26
[6]Cyranoski D,Reardon S.Embryo editing sparks epic debate.Nature,2015,520:593-4
[7]Ma H,Marti-Gutierrez N,Park SW,et al.Correction of a pathogenic gene mutation in human embryos.Nature,2017,548:413-9
[8]Pei D,Beier DW,Levy-Lahad E,et al.Human embryo editing:opportunities and importance of transnational cooperation.Cell Stem Cell,2017,21:423-6
[9]Vog G.Embryo edit makes human‘knockout’.Science,2017,357:1225
[10]Rossant J.Gene editing in human development:ethical concerns and practical applications.Development,2018,145,pii:dev150888
[11]Callaway E.Gene-editing research in human embryos gains momentum.Nature,2016,532:289-90
[12]Liang P,Xu Y,Zhang X,et al.CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes.Protein Cell,2015,6:363-72
[13]Ishii T.Germ line genome editing in clinics:the approaches,objectives and global society.Brief Funct Genomics,2017,16:46-56
[14]Li G,Liu Y,Zeng Y,et al.Highly efficient and precise base editing in discarded human tripronuclear embryos.Protein Cell,2017,8:776-9
[15]Kang XJ,He WY,Huang YL,et al.Introducing precise genetic modifications into human 3PN embryos by CRIS-PR/Cas-mediated genome editing.J Assist Reprod Genet,2016,33:581-8
[16]Fogarty NM,McCarthy A,Snijders KE,et al.Genome editing reveals a role for OCT4 in human embryogenesis.Nature,2017,550:67-73
[17]Tang L,Zeng Y,Du H,et al.CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein.Mol Genet Genomics,2017,292:525-33
[18]Ruzo A,Brivanlou AH.At last:gene editing in human embryos to understand human development.Cell Stem Cell,2017,21:564-5
[19]Cyranoski D.Ethics of embryo editing divides scientists.Nature,2015,519:272
[20]Baltimore D,Berg P,Botchan M,et al.Biotechnology.Aprudent path forward for genomic engineering and germline gene modification.Science,2015,348:36-8
[21]Martikainen M,Pedersen O.Germline edits:heat does not help debate.Nature,2015,520:623
[22]Savulescu J,Gyngell C,Douglas T.Germline edits:trust ethics review process.Nature,2015,520:623
[23]Porteus MH,Dann CT.Genome editing of the germline:broadening the discussion.Mol Ther,2015,23:980-2
[24]Friedmann T,Jonlin EC,King NMP,et al.ASGCT and JSGT joint position statement on human genomic editing.Mol Ther,2015,23:1282
[25]Lanphier E,Urnov F,Haecker SE,et al.Don’t edit the human germ line.Nature,2015,519:410-1
[26]Miller HI.Germline gene therapy:we’re ready.Science,2015,348:1325
[27]Lunshof JE.Human germ line editing-roles and responsibilities.Protein Cell,2016,7:7-10
[28]Addison C,Taylor-Alexander S.Gene editing and germline intervention:the need for novel responses to novel technologies.Mol Ther,2015,23:1678-80
[29]Thompson C.Human embryos:collect reliable data on embryo selection.Nature,2017,551:33
[30]Regalado A.Engineering the perfect baby.Technol Rev,2015,118:26-33
[31]Cartier-Lacave N,Ali R,Yla-Herttuala S,et al.Debate on germline gene editing.Hum Gene Ther Methods,2016,27:135-42
[32]Ishii T.Reproductive medicine involving genome editing:clinical uncertainties and embryological needs.Reprod Biomed Online,2017,34:27-31
[33]Savulescu J,Pugh J,Douglas T,et al.The moral imperative to continue gene editing research on human embryos.Protein Cell,2015,6:476-9
[34]Ball P.Kathy Niakan:at the forefront of gene editing in embryos.Lancet,2016,387:935-5
[35]Kaiser J.A yellow light for embryo editing.Science,2017,355:675
[36]范月蕾,王慧媛,于建荣.基因编辑的伦理争议.科技中国,2018:98-104
[37]Ormond KE,Mortlock DP,Scholes DT,et al.Human germline genome editing.Am J Hum Genet,2017,101:167-76
[38]Plaza Reyes A,Lanner F.Towards a CRISPR view of early human development:applications,limitations and ethical concerns of genome editing in human embryos.Development,2017,144:3-7
[39]Human embryo research policy update.Nat Biotechnol,2018,36:477
[40]Austin CP,Dawkins HJS.Medical research:next decade’s goals for rare diseases.Nature,2017,548:158
[41]Hsu PD,Lander ES,Zhang F.Development and applications of CRISPR-Cas9 for genome engineering.Cell,2014,157:1262-78
[42]Gross M.Bacterial scissors to edit human embryos?Curr Biol,2015,25:R439-42
[43]Ledford H.CRISPR fixes disease gene in viable human embryos.Nature,2017,548:13-4
[44]Egli D,Zuccaro MV,Kosicki M,et al.Inter-homologue repair in fertilized human eggs?Nature,2018,560:E5-E7
[45]Schenkwein D,Yla-Herttuala S.Gene editing of human embryos with CRISPR/Cas9:Great promise coupled with important caveats.Mol Ther,2018,26:659-60
[46]Kosicki M,Tomberg K,Bradley A.Repair of double-strand breaks induced by CRISPR-Cas9 leads to large deletions and complex rearrangements.Nat Biotechnol,2018,36:765-71
[47]Haapaniemi E,Botla S,Persson J,et al.CRISPR-Cas9genome editing induces a p53-mediated DNA damage response.Nat Med,2018,24:927-30
[48]Ihry RJ,Worringer KA,Salick MR,et al.p53 inhibits CRISPR-Cas9 engineering in human pluripotent stem cells.Nat Med,2018,24:939-46
[49]Eid A,Alshareef S,Mahfouz MM.CRISPR base editors:genome editing without double-stranded breaks.Biochem J,2018,475:1955-64
[50]Hess GT,Tycko J,Yao D,et al.Methods and applications of CRISPR-mediated base editing in eukaryotic genomes.Mol Cell,2017,68:26-43
[51]Komor AC,Kim YB,Packer MS,et al.Programmable editing of a target base in genomic DNA without doublestranded DNA cleavage.Nature,2016,533:420-4
[52]Kim D,Lim K,Kim ST,et al.Genome-wide target specificities of CRISPR RNA-guided programmable deaminases.Nat Biotechnol,2017,35:475-80
[53]Koblan LW,Doman JL,Wilson C,et al.Improving cytidine and adenine base editors by expression optimization and ancestral reconstruction.Nat Biotechnol,2018,36:843-6
[54]Li X,Wang Y,Liu Y,et al.Base editing with a Cpf1-cytidine deaminase fusion.Nat Biotechnol,2018,36:324-7
[55]Wang X,Li J,Wang Y,et al.Efficient base editing in methylated regions with a human APOBEC3A-Cas9fusion.Nat Biotechnol,2018[Epub ahead of print]
[56]Kim YB,Komor AC,Levy JM,et al.Increasing the genome-targeting scope and precision of base editing with engineered Cas9-cytidine deaminase fusions.Nat Biotechnol,2017,35:371-6
[57]Jiang W,Feng S,Huang S,et al.BE-PLUS:a new base editing tool with broadened editing window and enhanced fidelity.Cell Res,2018[Epub ahead of print]
[58]Gaudelli NM,Komor AC,Rees HA,et al.Programmable base editing of A*T to G*C in genomic DNA without DNA cleavage.Nature,2017,551:464-71
[59]Liu Z,Chen M,Chen S,et al.Highly efficient RNA-guided base editing in rabbit.Nat Commun,2018,9:2717
[60]Li C,Zong Y,Wang Y,et al.Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion.Genome Biol,2018,19:59
[61]Liu Z,Lu Z,Yang G,et al.Efficient generation of mouse models of human diseases via ABE-and BE-mediated base editing.Nat Commun,2018,9:2338
[62]Zhou C,Zhang M,Wei Y,et al.Highly efficient base editing in human tripronuclear zygotes.Protein Cell,2017,8:772-5
[63]Liang PP,Ding CH,Sun HW,et al.Correction ofβ-thalassemia mutant by base editor in human embryos.Protein Cell,2017,8:811-22
[64]Zeng Y,Li J,Li G,et al.Correction of the Marfan syndrome pathogenic FBN1 mutation by base editing in human cells and heterozygous embryos.Mol Therapy,2018.pii:S1525-0016(18)30378-2
[65]Aird EJ,Lovendahl KN,St.Martin A,et al.Increasing Cas9-mediated homology-directed repair efficiency through covalent tethering of DNA repair template.Commun Biol,2018,1:54