Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing
Author(s)Kumita, Wakako; Sato, Kenya; Suzuki, Yasuhiro; Kurotaki, Yoko; Harada, Takeshi; Zhou, Yang; Kishi, Noriyuki; Sato, Kengo; Aiba, Atsu; Sakakibara, Yasubumi; Feng, Guoping; Okano, Hideyuki; Sasaki, Erika; ... Show more Show less
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Genetically modified nonhuman primates (NHP) are useful models for biomedical research. Gene editing technologies have enabled production of target-gene knock-out (KO) NHP models. Target-gene-KO/knock-in (KI) efficiency of CRISPR/Cas9 has not been extensively investigated in marmosets. In this study, optimum conditions for target gene modification efficacies of CRISPR/mRNA and CRISPR/nuclease in marmoset embryos were examined. CRISPR/nuclease was more effective than CRISPR/mRNA in avoiding mosaic genetic alteration. Furthermore, optimal conditions to generate KI marmoset embryos were investigated using CRISPR/Cas9 and 2 different lengths (36 nt and 100 nt) each of a sense or anti-sense single-strand oligonucleotide (ssODN). KIs were observed when CRISPR/nuclease and 36 nt sense or anti-sense ssODNs were injected into embryos. All embryos exhibited mosaic mutations with KI and KO, or imprecise KI, of c-kit. Although further improvement of KI strategies is required, these results indicated that CRISPR/Cas9 may be utilized to produce KO/KI marmosets via gene editing.
DepartmentMcGovern Institute for Brain Research at MIT; Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Springer Science and Business Media LLC
Kumita, Wakako et al. Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing. Sci Rep 9, 1 (September 2019): 12719 © 2019 The Author(s)
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