Efficient biallelic mutation in porcine parthenotes using a CRISPR-Cas9 system
作者： Li Tao , Mingyao Yang , Xiaodong Wang , Zhenni Zhang , Zhonghong Wu
DOI: 10.1016/J.BBRC.2016.05.100
关键词:
摘要: The parthenotes represent ideal models mimicking the embryonic development and characterizing function of maternal genomes as well an alternative source pluripotent cell lines. Besides, parthenogenetically activated (PA) embryos serve a rapid assay system to maximize efficiency generating genetically modified pig CRISPR/Cas9 system, efficient multiplex gene editing tool, has been utilized modify genome porcine parthenotes. However, lower biallelic mutation rate high mosaicism frequency were observed. Here, we aimed enhance with reduced by optimization concentration injection time Cas9/sgRNA mixture in results showed that (93%) low (33%) could be achieved cytoplasmic Cas9 mRNA/sgRNA (125/12.5 ng/μl) after 8 h parthenogenetical activation. Thus, our study provides effective strategy for increasing population homogeneity parthenotes, which will strengthen role uncovering early assessing due simplicity adaptability CRISPR/Cas9.
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