Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing.

Autor: Kumita W; Central Institute for Experimental Animals, Kawasaki-shi, Kanagawa, 210-0821, Japan., Sato K; Central Institute for Experimental Animals, Kawasaki-shi, Kanagawa, 210-0821, Japan., Suzuki Y; Central Institute for Experimental Animals, Kawasaki-shi, Kanagawa, 210-0821, Japan., Kurotaki Y; Central Institute for Experimental Animals, Kawasaki-shi, Kanagawa, 210-0821, Japan., Harada T; Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Osaka University, Suita-shi, Osaka, 565-0871, Japan.; Laboratory of Animal Resources, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan., Zhou Y; McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, 02139, USA., Kishi N; Laboratory for Marmoset Neural Architecture, RIKEN Center for Brain Science, Wako-shi, Saitama, 351-0198, Japan., Sato K; Department of Biosciences and Informatics, Keio University, Yokohama-shi, Kanagawa, 223-8522, Japan., Aiba A; Laboratory of Animal Resources, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan., Sakakibara Y; Department of Biosciences and Informatics, Keio University, Yokohama-shi, Kanagawa, 223-8522, Japan., Feng G; McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, Massachusetts, 02139, USA., Okano H; Laboratory for Marmoset Neural Architecture, RIKEN Center for Brain Science, Wako-shi, Saitama, 351-0198, Japan.; Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, 160-8582, Japan., Sasaki E; Central Institute for Experimental Animals, Kawasaki-shi, Kanagawa, 210-0821, Japan. esasaki@ciea.or.jp.; Laboratory for Marmoset Neural Architecture, RIKEN Center for Brain Science, Wako-shi, Saitama, 351-0198, Japan. esasaki@ciea.or.jp.; Advanced Research Center, Keio University, Shinjuku-ku, Tokyo, 160-8582, Japan. esasaki@ciea.or.jp.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2019 Sep 03; Vol. 9 (1), pp. 12719. Date of Electronic Publication: 2019 Sep 03.
DOI: 10.1038/s41598-019-49110-3
Abstrakt: 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.
Databáze: MEDLINE
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