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CRISPR/Cas9-mediated generation of a Plac8 knockout mouse model

Laboratory Animal Research volume 34, pages 279–287 (2018)Cite this article

Abstract

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.

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  1. These authors contributed equally to this work

Authors and Affiliations

  1. Department of Biochemistry, College of Life Science & Biotechnology, Yonsei University, Seoul, 03722, Korea

    HyunJeong Lee, Jae-il Roh, Jaehoon Lee & Han-Woong Lee

  2. Graduate School of Translational Medicine, Seoul National University College of Medicine, 101 Daehakro, Jongno-gu, Seoul, 110-744, Korea

    Joo-Il Kim & Byeong-Cheol Kang

  3. Department of Experimental Animal Research, Biomedical Research Institute, Seoul National Univ. Hospital, Seoul, Korea

    Joo-Il Kim, Jin-Sung Park & Byeong-Cheol Kang

  4. Designed Animal and Transplantation Research Institute, Institute of GreenBio Science Technology, Seoul National University, Pyeongchang-gun, Korea

    Byeong-Cheol Kang

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  1. HyunJeong Lee
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Correspondence to Jae-il Roh, Jaehoon Lee, Byeong-Cheol Kang or Han-Woong Lee.

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Lee, H., Kim, JI., Park, JS. et al. CRISPR/Cas9-mediated generation of a Plac8 knockout mouse model. Lab Anim Res 34, 279–287 (2018). https://doi.org/10.5625/lar.2018.34.4.279

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  • DOI: https://doi.org/10.5625/lar.2018.34.4.279

Keywords

Laboratory Animal Research

ISSN: 2233-7660