文章基本信息

标题：CRISPR/Cas9-mediated generation of a Plac8 knockout mouse model
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作者：HyunJeong Lee ; Joo-Il Kim ; Jin-Sung Park 等
期刊名称：Laboratory Animal Research
印刷版ISSN：1738-6055
电子版ISSN：2233-7660
出版年度：2018
卷号：34
期号：4
页码：279-287
DOI：10.5625/lar.2018.34.4.279
语种：English
出版社：BioMed Central Ltd.
摘要：Placenta specific 8 (PLAC8, also known as ONZIN) is a multi-functional protein that is highly expressed in the intestine, lung, spleen, and innate immune cells, and is involved in various diseases, including cancers, obesity, and innate immune deficiency. Here, we generated a Plac8 knockout mouse using the CRISPR/Cas9 system. The Cas9 mRNA and two single guide RNAs targeting a region near the translation start codon at Plac8 exon 2 were microinjected into mouse zygotes. This successfully eliminated the conventional translation start site, as confirmed by Sanger sequencing and PCR genotyping analysis. Unlike the previous Plac8 deficient models displaying increased adipose tissue and body weights, our male Plac8 knockout mice showed rather lower body weight than sex-matched littermate controls, though the only difference between these two mouse models is genetic context. Differently from the previously constructed embryonic stem cell-derived Plac8 knockout mouse that contains a neomycin resistance cassette, this knockout mouse model is free from a negative selection marker or other external insertions, which will be useful in future studies aimed at elucidating the multi-functional and physiological roles of PLAC8 in various diseases, without interference from exogenous foreign DNA.
关键词：PLAC8;CRISPR/Cas9;mouse model