CRISPR/Cas9-mediated generation of a Plac8 knockout mouse model.
10.5625/lar.2018.34.4.279
- Author:
HyunJeong LEE
1
;
Joo Il KIM
;
Jin Sung PARK
;
Jae il ROH
;
Jaehoon LEE
;
Byeong Cheol KANG
;
Han Woong LEE
Author Information
1. Department of Biochemistry, College of Life Science and Biotechnology and Yonsei Laboratory Animal Research Center, Yonsei University, Seoul, Korea. hwl@yonsei.ac.kr, jhlee13@gmail.com, rohjaeil@gmail.com
- Publication Type:Original Article
- Keywords:
PLAC8;
CRISPR/Cas9;
mouse model
- MeSH:
Adipose Tissue;
Animals;
Body Weight;
Codon, Initiator;
DNA;
Exons;
Humans;
Intestines;
Lung;
Male;
Mice;
Mice, Knockout*;
Neomycin;
Obesity;
Placenta;
Polymerase Chain Reaction;
RNA, Guide;
RNA, Messenger;
Spleen;
Zygote
- From:Laboratory Animal Research
2018;34(4):279-287
- CountryRepublic of Korea
- Language:English
-
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.
