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Stem Cells International
Volume 2018 (2018), Article ID 4136473, 11 pages
https://doi.org/10.1155/2018/4136473
Review Article

Genome Editing Redefines Precision Medicine in the Cardiovascular Field

1Department of Cardiovascular Surgery, German Heart Center Munich, Technische Universität München, Lazarettstraße 36, 80636 Munich, Germany
2Insure (Institute for Translational Cardiac Surgery), Department of Cardiovascular Surgery, German Heart Center, Technische Universität München, Lothstraße 11, 80636 Munich, Germany
3German Center for Cardiovascular Research (DZHK), Partner Site Munich Heart Alliance, Munich, Germany
4Department of Medicine, Division of Cardiovascular Medicine, Cardiovascular Institute, and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, 265 Campus Drive, Stanford, CA 94305, USA

Correspondence should be addressed to Elda Dzilic; ed.nhm.mhd@cilizd and Stefanie A. Doppler; moc.liamg@relppodiffets

Received 5 July 2017; Accepted 25 October 2017; Published 14 March 2018

Academic Editor: Andrzej Lange

Copyright © 2018 Elda Dzilic et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Linked References

  1. E. S. Lander, L. M. Linton, B. Birren et al., “Initial sequencing and analysis of the human genome,” Nature, vol. 409, no. 6822, pp. 860–921, 2001. View at Publisher · View at Google Scholar · View at Scopus
  2. J. Travis, “Cover stories: making the breakthrough of the year cover,” Science, vol. 354, no. 6319, p. 1497, 2016. View at Publisher · View at Google Scholar · View at Scopus
  3. T. Doetschman and T. Georgieva, “Gene editing with CRISPR/Cas9 RNA-directed nuclease,” Circulation Research, vol. 120, no. 5, pp. 876–894, 2017. View at Publisher · View at Google Scholar · View at Scopus
  4. J. W. Buikema and S. M. Wu, “Untangling the biology of genetic cardiomyopathies with pluripotent stem cell disease models,” Current Cardiology Reports, vol. 19, no. 4, p. 30, 2017. View at Publisher · View at Google Scholar · View at Scopus
  5. N. Brookhouser, S. Raman, C. Potts, and D. Brafman, “May I cut in? Gene editing approaches in human induced pluripotent stem cells,” Cells, vol. 6, no. 4, p. 5, 2017. View at Publisher · View at Google Scholar
  6. D. Waldron, “Gene therapy: in vivo gene editing in non-dividing cells,” Nature Reviews Genetics, vol. 18, no. 1, p. 1, 2017. View at Publisher · View at Google Scholar · View at Scopus
  7. T. Ishizu, S. Higo, Y. Masumura et al., “Targeted genome replacement via homology-directed repair in non-dividing cardiomyocytes,” Scientific Reports, vol. 7, no. 1, p. 9363, 2017. View at Publisher · View at Google Scholar · View at Scopus
  8. K. Takahashi, K. Tanabe, M. Ohnuki et al., “Induction of pluripotent stem cells from adult human fibroblasts by defined factors,” Cell, vol. 131, no. 5, pp. 861–872, 2007. View at Publisher · View at Google Scholar · View at Scopus
  9. K. Takahashi and S. Yamanaka, “Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors,” Cell, vol. 126, no. 4, pp. 663–676, 2006. View at Publisher · View at Google Scholar · View at Scopus
  10. G. Wang, M. L. McCain, L. Yang et al., “Modeling the mitochondrial cardiomyopathy of Barth syndrome with induced pluripotent stem cell and heart-on-chip technologies,” Nature Medicine, vol. 20, no. 6, pp. 616–623, 2014. View at Publisher · View at Google Scholar · View at Scopus
  11. I. Karakikes, F. Stillitano, M. Nonnenmacher et al., “Correction of human phospholamban R14del mutation associated with cardiomyopathy using targeted nucleases and combination therapy,” Nature Communications, vol. 6, no. 1, p. 6955, 2015. View at Publisher · View at Google Scholar · View at Scopus
  12. C. V. Theodoris, M. Li, M. P. White et al., “Human disease modeling reveals integrated transcriptional and epigenetic mechanisms of NOTCH1 haploinsufficiency,” Cell, vol. 160, no. 6, pp. 1072–1086, 2015. View at Publisher · View at Google Scholar · View at Scopus
  13. J. T. Hinson, A. Chopra, N. Nafissi et al., “Titin mutations in iPS cells define sarcomere insufficiency as a cause of dilated cardiomyopathy,” Science, vol. 349, no. 6251, pp. 982–986, 2015. View at Publisher · View at Google Scholar · View at Scopus
  14. K. Kodo, S. G. Ong, F. Jahanbani et al., “iPSC-derived cardiomyocytes reveal abnormal TGF-β signalling in left ventricular non-compaction cardiomyopathy,” Nature Cell Biology, vol. 18, no. 10, pp. 1031–1042, 2016. View at Publisher · View at Google Scholar · View at Scopus
  15. Y. Yamamoto, T. Makiyama, T. Harita et al., “Allele-specific ablation rescues electrophysiological abnormalities in a human iPS cell model of long-QT syndrome with a CALM2 mutation,” Human Molecular Genetics, vol. 26, no. 9, pp. 1670–1677, 2017. View at Publisher · View at Google Scholar · View at Scopus
  16. M. Bellin, S. Casini, R. P. Davis et al., “Isogenic human pluripotent stem cell pairs reveal the role of a KCNH2 mutation in long-QT syndrome,” The EMBO Journal, vol. 32, no. 24, pp. 3161–3175, 2013. View at Publisher · View at Google Scholar · View at Scopus
  17. R. M. Gupta, T. B. Meissner, C. A. Cowan, and K. Musunuru, “Genome-edited human pluripotent stem cell–derived macrophages as a model of reverse cholesterol transport—brief report,” Arteriosclerosis, Thrombosis, and Vascular Biology, vol. 36, no. 1, pp. 15–18, 2016. View at Publisher · View at Google Scholar · View at Scopus
  18. I. Karakikes, V. Termglinchan, D. A. Cepeda et al., “A comprehensive TALEN-based knockout library for generating human-induced pluripotent stem cell–based models for cardiovascular diseases,” Circulation Research, vol. 120, no. 10, pp. 1561–1571, 2017. View at Publisher · View at Google Scholar · View at Scopus
  19. Y. Wang, P. Liang, F. Lan et al., “Genome editing of isogenic human induced pluripotent stem cells recapitulates long QT phenotype for drug testing,” Journal of the American College of Cardiology, vol. 64, no. 5, pp. 451–459, 2014. View at Publisher · View at Google Scholar · View at Scopus
  20. C. Robertson, D. D. Tran, and S. C. George, “Concise review: maturation phases of human pluripotent stem cell-derived cardiomyocytes,” Stem Cells, vol. 31, no. 5, pp. 829–837, 2013. View at Publisher · View at Google Scholar · View at Scopus
  21. T. J. Kolanowski, C. L. Antos, and K. Guan, “Making human cardiomyocytes up to date: derivation, maturation state and perspectives,” International Journal of Cardiology, vol. 241, pp. 379–386, 2017. View at Publisher · View at Google Scholar · View at Scopus
  22. R. Jha, B. Wile, Q. Wu et al., “Molecular beacon-based detection and isolation of working-type cardiomyocytes derived from human pluripotent stem cells,” Biomaterials, vol. 50, pp. 176–185, 2015. View at Publisher · View at Google Scholar · View at Scopus
  23. A. M. Wiencierz, M. Kernbach, J. Ecklebe et al., “Differential expression levels of integrin α6 enable the selective identification and isolation of atrial and ventricular cardiomyocytes,” PLoS One, vol. 10, no. 11, article e0143538, 2015. View at Publisher · View at Google Scholar · View at Scopus
  24. Z. Chen, W. Xian, M. Bellin et al., “Subtype-specific promoter-driven action potential imaging for precise disease modelling and drug testing in hiPSC-derived cardiomyocytes,” European Heart Journal, vol. 38, no. 4, pp. 292–301, 2016. View at Publisher · View at Google Scholar · View at Scopus
  25. A. Bizy, G. Guerrero-Serna, B. Hu et al., “Myosin light chain 2-based selection of human iPSC-derived early ventricular cardiac myocytes,” Stem Cell Research, vol. 11, no. 3, pp. 1335–1347, 2013. View at Publisher · View at Google Scholar · View at Scopus
  26. R. Josowitz, J. Lu, C. Falce et al., “Identification and purification of human induced pluripotent stem cell-derived atrial-like cardiomyocytes based on sarcolipin expression,” PLoS One, vol. 9, no. 7, article e101316, 2014. View at Publisher · View at Google Scholar · View at Scopus
  27. L. A. Gilbert, M. A. Horlbeck, B. Adamson et al., “Genome-scale CRISPR-mediated control of gene repression and activation,” Cell, vol. 159, no. 3, pp. 647–661, 2014. View at Publisher · View at Google Scholar · View at Scopus
  28. L. S. Qi, M. H. Larson, L. A. Gilbert et al., “Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression,” Cell, vol. 152, no. 5, pp. 1173–1183, 2013. View at Publisher · View at Google Scholar · View at Scopus
  29. A. L. Jackson, S. R. Bartz, J. Schelter et al., “Expression profiling reveals off-target gene regulation by RNAi,” Nature Biotechnology, vol. 21, no. 6, pp. 635–637, 2003. View at Publisher · View at Google Scholar · View at Scopus
  30. S. Qiu, C. M. Adema, and T. Lane, “A computational study of off-target effects of RNA interference,” Nucleic Acids Research, vol. 33, no. 6, pp. 1834–1847, 2005. View at Publisher · View at Google Scholar · View at Scopus
  31. F. Zhang, L. Cong, S. Lodato, S. Kosuri, G. M. Church, and P. Arlotta, “Efficient construction of sequence-specific TAL effectors for modulating mammalian transcription,” Nature Biotechnology, vol. 29, no. 2, pp. 149–153, 2011. View at Publisher · View at Google Scholar · View at Scopus
  32. P. Mali, J. Aach, P. B. Stranges et al., “CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering,” Nature Biotechnology, vol. 31, no. 9, pp. 833–838, 2013. View at Publisher · View at Google Scholar · View at Scopus
  33. P. Perez-Pinera, D. D. Kocak, C. M. Vockley et al., “RNA-guided gene activation by CRISPR-Cas9–based transcription factors,” Nature Methods, vol. 10, no. 10, pp. 973–976, 2013. View at Publisher · View at Google Scholar · View at Scopus
  34. X. Gao, J. Yang, J. C. H. Tsang, J. Ooi, D. Wu, and P. Liu, “Reprogramming to pluripotency using designer TALE transcription factors targeting enhancers,” Stem Cell Reports, vol. 1, no. 2, pp. 183–197, 2013. View at Publisher · View at Google Scholar · View at Scopus
  35. S. Bultmann, R. Morbitzer, C. S. Schmidt et al., “Targeted transcriptional activation of silent oct4 pluripotency gene by combining designer TALEs and inhibition of epigenetic modifiers,” Nucleic Acids Research, vol. 40, no. 12, pp. 5368–5377, 2012. View at Publisher · View at Google Scholar · View at Scopus
  36. A. W. Cheng, H. Wang, H. Yang et al., “Multiplexed activation of endogenous genes by CRISPR-on, an RNA-guided transcriptional activator system,” Cell Research, vol. 23, no. 10, pp. 1163–1171, 2013. View at Publisher · View at Google Scholar · View at Scopus
  37. J. Hu, Y. Lei, W. K. Wong et al., “Direct activation of human and mouse Oct4 genes using engineered TALE and Cas9 transcription factors,” Nucleic Acids Research, vol. 42, no. 7, pp. 4375–4390, 2014. View at Publisher · View at Google Scholar · View at Scopus
  38. N. Fusaki, H. Ban, A. Nishiyama, K. Saeki, and M. Hasegawa, “Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome,” Proceedings of the Japan Academy, Series B, vol. 85, no. 8, pp. 348–362, 2009. View at Publisher · View at Google Scholar · View at Scopus
  39. S. Chakraborty, H. Y. Ji, A. M. Kabadi, C. A. Gersbach, N. Christoforou, and K. W. Leong, “A CRISPR/Cas9-based system for reprogramming cell lineage specification,” Stem Cell Reports, vol. 3, no. 6, pp. 940–947, 2014. View at Publisher · View at Google Scholar · View at Scopus
  40. C. Mason and P. Dunnill, “A brief definition of regenerative medicine,” Regenerative Medicine, vol. 3, no. 1, pp. 1–5, 2008. View at Publisher · View at Google Scholar · View at Scopus
  41. P. Macchiarini, P. Jungebluth, T. Go et al., “Clinical transplantation of a tissue-engineered airway,” The Lancet, vol. 372, no. 9655, pp. 2023–2030, 2008. View at Publisher · View at Google Scholar · View at Scopus
  42. P. Menasche, V. Vanneaux, J.-R. Fabreguettes et al., “Towards a clinical use of human embryonic stem cell-derived cardiac progenitors: a translational experience,” European Heart Journal, vol. 36, no. 12, pp. 743–750, 2015. View at Publisher · View at Google Scholar · View at Scopus
  43. J. J. H. Chong, X. Yang, C. W. Don et al., “Human embryonic-stem-cell-derived cardiomyocytes regenerate non-human primate hearts,” Nature, vol. 510, no. 7504, pp. 273–277, 2014. View at Publisher · View at Google Scholar · View at Scopus
  44. V. Lepperhof, O. Polchynski, K. Kruttwig et al., “Bioluminescent imaging of genetically selected induced pluripotent stem cell-derived cardiomyocytes after transplantation into infarcted heart of syngeneic recipients,” PLoS One, vol. 9, no. 9, article e107363, 2014. View at Publisher · View at Google Scholar · View at Scopus
  45. S. V. Rojas, G. Kensah, A. Rotaermel et al., “Transplantation of purified iPSC-derived cardiomyocytes in myocardial infarction,” PLoS One, vol. 12, no. 5, article e0173222, 2017. View at Publisher · View at Google Scholar · View at Scopus
  46. Y. Shiba, T. Gomibuchi, T. Seto et al., “Allogeneic transplantation of iPS cell-derived cardiomyocytes regenerates primate hearts,” Nature, vol. 538, no. 7625, pp. 388–391, 2016. View at Publisher · View at Google Scholar · View at Scopus
  47. T. Freyman, G. Polin, H. Osman et al., “A quantitative, randomized study evaluating three methods of mesenchymal stem cell delivery following myocardial infarction,” European Heart Journal, vol. 27, no. 9, pp. 1114–1122, 2006. View at Publisher · View at Google Scholar · View at Scopus
  48. P. Menasché, V. Vanneaux, A. Hagège et al., “Human embryonic stem cell-derived cardiac progenitors for severe heart failure treatment: first clinical case report,” European Heart Journal, vol. 36, no. 30, pp. 2011–2017, 2015. View at Publisher · View at Google Scholar · View at Scopus
  49. K. Suzuki, Y. Tsunekawa, R. Hernandez-Benitez et al., “In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration,” Nature, vol. 540, no. 7631, pp. 144–149, 2016. View at Publisher · View at Google Scholar · View at Scopus
  50. M. Gramlich, L. S. Pane, Q. Zhou et al., “Antisense-mediated exon skipping: a therapeutic strategy for titin-based dilated cardiomyopathy,” EMBO Molecular Medicine, vol. 7, no. 5, pp. 562–576, 2015. View at Publisher · View at Google Scholar · View at Scopus
  51. C. Gedicke-Hornung, V. Behrens-Gawlik, S. Reischmann et al., “Rescue of cardiomyopathy through U7snRNA-mediated exon skipping in Mybpc3-targeted knock-in mice,” EMBO Molecular Medicine, vol. 5, no. 7, pp. 1128–1145, 2013. View at Publisher · View at Google Scholar · View at Scopus
  52. C. Long, L. Amoasii, A. A. Mireault et al., “Postnatal genome editing partially restores dystrophin expression in a mouse model of muscular dystrophy,” Science, vol. 351, no. 6271, pp. 400–403, 2016. View at Publisher · View at Google Scholar · View at Scopus
  53. C. E. Nelson, C. H. Hakim, D. G. Ousterout et al., “In vivo genome editing improves muscle function in a mouse model of Duchenne muscular dystrophy,” Science, vol. 351, no. 6271, pp. 403–407, 2016. View at Publisher · View at Google Scholar · View at Scopus
  54. M. Tabebordbar, K. Zhu, J. K. W. Cheng et al., “In vivo gene editing in dystrophic mouse muscle and muscle stem cells,” Science, vol. 351, no. 6271, pp. 407–411, 2016. View at Publisher · View at Google Scholar · View at Scopus
  55. Q. Ding, A. Strong, K. M. Patel et al., “Permanent alteration of PCSK9 with in vivo CRISPR-Cas9 genome editing,” Circulation Research, vol. 115, no. 5, pp. 488–492, 2014. View at Publisher · View at Google Scholar · View at Scopus
  56. J. C. Cohen, E. Boerwinkle, T. H. Mosley Jr., and H. H. Hobbs, “Sequence variations in PCSK9, low LDL, and protection against coronary heart disease,” The New England Journal of Medicine, vol. 354, no. 12, pp. 1264–1272, 2006. View at Publisher · View at Google Scholar · View at Scopus
  57. S. W. M. van den Borne, J. Diez, W. M. Blankesteijn, J. Verjans, L. Hofstra, and J. Narula, “Myocardial remodeling after infarction: the role of myofibroblasts,” Nature Reviews Cardiology, vol. 7, no. 1, pp. 30–37, 2010. View at Publisher · View at Google Scholar · View at Scopus
  58. P. Liang, Y. Xu, X. Zhang et al., “CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes,” Protein & Cell, vol. 6, no. 5, pp. 363–372, 2015. View at Publisher · View at Google Scholar · View at Scopus
  59. L. Tang, Y. Zeng, H. du et al., “CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein,” Molecular Genetics and Genomics, vol. 292, no. 3, pp. 525–533, 2017. View at Publisher · View at Google Scholar · View at Scopus
  60. H. Ma, N. Marti-Gutierrez, S. W. Park et al., “Correction of a pathogenic gene mutation in human embryos,” Nature, vol. 548, no. 7668, pp. 413–419, 2017. View at Publisher · View at Google Scholar · View at Scopus
  61. S. Q. Tsai, Z. Zheng, N. T. Nguyen et al., “GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases,” Nature Biotechnology, vol. 33, no. 2, pp. 187–197, 2015. View at Publisher · View at Google Scholar · View at Scopus
  62. T. J. Cradick, E. J. Fine, C. J. Antico, and G. Bao, “CRISPR/Cas9 systems targeting β-globin and CCR5 genes have substantial off-target activity,” Nucleic Acids Research, vol. 41, no. 20, pp. 9584–9592, 2013. View at Publisher · View at Google Scholar · View at Scopus
  63. Y. Fu, J. A. Foden, C. Khayter et al., “High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells,” Nature Biotechnology, vol. 31, no. 9, pp. 822–826, 2013. View at Publisher · View at Google Scholar · View at Scopus
  64. R. L. Frock, J. Hu, R. M. Meyers, Y. J. Ho, E. Kii, and F. W. Alt, “Genome-wide detection of DNA double-stranded breaks induced by engineered nucleases,” Nature Biotechnology, vol. 33, no. 2, pp. 179–186, 2015. View at Publisher · View at Google Scholar · View at Scopus
  65. G. Schwank, B. K. Koo, V. Sasselli et al., “Functional repair of CFTR by CRISPR/Cas9 in intestinal stem cell organoids of cystic fibrosis patients,” Cell Stem Cell, vol. 13, no. 6, pp. 653–658, 2013. View at Publisher · View at Google Scholar · View at Scopus
  66. U. Weissbein, N. Benvenisty, and U. Ben-David, “Quality control: genome maintenance in pluripotent stem cells,” The Journal of Cell Biology, vol. 204, no. 2, pp. 153–163, 2014. View at Publisher · View at Google Scholar · View at Scopus
  67. C. Smith, A. Gore, W. Yan et al., “Whole-genome sequencing analysis reveals high specificity of CRISPR/Cas9 and TALEN-based genome editing in human iPSCs,” Cell Stem Cell, vol. 15, no. 1, pp. 12-13, 2014. View at Publisher · View at Google Scholar · View at Scopus
  68. H. Wang, H. Yang, C. S. Shivalila et al., “One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering,” Cell, vol. 153, no. 4, pp. 910–918, 2013. View at Publisher · View at Google Scholar · View at Scopus
  69. Y. Niu, B. Shen, Y. Cui et al., “Generation of gene-modified cynomolgus monkey via Cas9/RNA-mediated gene targeting in one-cell embryos,” Cell, vol. 156, no. 4, pp. 836–843, 2014. View at Publisher · View at Google Scholar · View at Scopus
  70. J. Mianné, L. Chessum, S. Kumar et al., “Correction of the auditory phenotype in C57BL/6N mice via CRISPR/Cas9-mediated homology directed repair,” Genome Medicine, vol. 8, no. 1, p. 16, 2016. View at Publisher · View at Google Scholar · View at Scopus
  71. Y. Fu, J. D. Sander, D. Reyon, V. M. Cascio, and J. K. Joung, “Improving CRISPR-Cas nuclease specificity using truncated guide RNAs,” Nature Biotechnology, vol. 32, no. 3, pp. 279–284, 2014. View at Publisher · View at Google Scholar · View at Scopus
  72. B. P. Kleinstiver, V. Pattanayak, M. S. Prew et al., “High–fidelity CRISPR–Cas9 nucleases with no detectable genome–wide off–target effects,” Nature, vol. 529, no. 7587, pp. 490–495, 2016. View at Publisher · View at Google Scholar · View at Scopus
  73. B. P. Kleinstiver, M. S. Prew, S. Q. Tsai et al., “Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition,” Nature Biotechnology, vol. 33, no. 12, pp. 1293–1298, 2015. View at Publisher · View at Google Scholar · View at Scopus
  74. B. P. Kleinstiver, M. S. Prew, S. Q. Tsai et al., “Engineered CRISPR-Cas9 nucleases with altered PAM specificities,” Nature, vol. 523, no. 7561, pp. 481–485, 2015. View at Publisher · View at Google Scholar · View at Scopus
  75. F. A. Ran, P. D. Hsu, C. Y. Lin et al., “Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity,” Cell, vol. 154, no. 6, pp. 1380–1389, 2013. View at Publisher · View at Google Scholar · View at Scopus
  76. Y. Wu, T. Gao, X. Wang et al., “TALE nickase mediates high efficient targeted transgene integration at the human multi-copy ribosomal DNA locus,” Biochemical and Biophysical Research Communications, vol. 446, no. 1, pp. 261–266, 2014. View at Publisher · View at Google Scholar · View at Scopus
  77. J. Wang, G. Friedman, Y. Doyon et al., “Targeted gene addition to a predetermined site in the human genome using a ZFN-based nicking enzyme,” Genome Research, vol. 22, no. 7, pp. 1316–1326, 2012. View at Publisher · View at Google Scholar · View at Scopus
  78. A. C. Komor, Y. B. Kim, M. S. Packer, J. A. Zuris, and D. R. Liu, “Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage,” Nature, vol. 533, no. 7603, pp. 420–424, 2016. View at Publisher · View at Google Scholar · View at Scopus
  79. N. M. Gaudelli, A. C. Komor, H. A. Rees et al., “Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage,” Nature, vol. 551, no. 7681, pp. 464–471, 2017. View at Publisher · View at Google Scholar · View at Scopus
  80. Y. B. Kim, A. C. Komor, J. M. Levy, M. S. Packer, K. T. Zhao, and D. R. Liu, “Increasing the genome-targeting scope and precision of base editing with engineered Cas9-cytidine deaminase fusions,” Nature Biotechnology, vol. 35, no. 4, pp. 371–376, 2017. View at Publisher · View at Google Scholar · View at Scopus
  81. C. D. Richardson, G. J. Ray, M. A. DeWitt, G. L. Curie, and J. E. Corn, “Enhancing homology-directed genome editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA,” Nature Biotechnology, vol. 34, no. 3, pp. 339–344, 2016. View at Publisher · View at Google Scholar · View at Scopus