Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing
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s41598-019-49110-3.pdf
Description
Published version
Size
2.51 MB
Format
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Checksum (MD5)
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Author(s)
Kumita, Wakako
Sato, Kenya
Suzuki, Yasuhiro
Kurotaki, Yoko
Harada, Takeshi
Zhou, Yang
Kishi, Noriyuki
Sato, Kengo
Aiba, Atsu
Sakakibara, Yasubumi
Date Issued
September 2019
Journal
Scientific Reports
Publisher
Springer Science and Business Media LLC
Citation
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)
Version
Final published version
Abstract
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.
MIT Department
McGovern Institute for Brain Research at MIT
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Terms of Use
Creative Commons Attribution 4.0 International license
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DOI of Published Version
https://doi.org/10.1038/s41598-019-49110-3
