Establishment of MrgD knockout mice model with CRISPR/Cas9 gene targeting technology

doi:10.3969/j.issn.1006-7795.2018.04.007

Journal of Capital Medical University ›› 2018, Vol. 39 ›› Issue (4): 517-521.doi: 10.3969/j.issn.1006-7795.2018.04.007

Previous Articles Next Articles

Establishment of MrgD knockout mice model with CRISPR/Cas9 gene targeting technology

Cao Xi^1,2, Song Lini^1,2, Zhang Yichen^1,2, Li Qi^1,2, Yang Jinkui^1,2

1. Department of Endocrinology, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China;
2. Beijing Key Laboratory of Diabetes Research and Care, Beijing 100730, China

Received:2018-06-04 Online:2018-07-21 Published:2018-07-21
Supported by:
This study was supported by National Natural Science Foundation of China (81670774, 81471014), Natural Science Foundation of Beijing(7162047).

Abstract

Abstract: Objective To establish the MrgD gene knockout mice model by using CRISPR/Cas9 gene editing technique so as to provide the tool to study the regulating role of MrgD in glucose and lipid metabolism. Methods Single guide RNA (sgRNA) plasmids against the exon 1 of MrgD were designed and constructed. Then the sgRNA and Cas9 was transcribed by T7 RNA polymerase in vitro. Cas9 mRNA and sgRNA were mixed and microinjected into fertilized eggs of C57BL/6J mice. Gene sequencing was used to detect the mutations of MrgD. Results Finally the F2 generation of homozygous MrgD deficient mice (MrgD-173/-173) were gained. Preliminary studies showed that MrgD gene homozygous mutant mice may not have disorders of glucose and lipid metabolism. ConclusionA mouse model with MrgD deficiency has been successfully established which shall lay a foundation for future investigation of MrgD.

Key words: gene knockout, CRISPR/Cas9, MrgD gene

CLC Number:

Q75
R587.1

Cao Xi, Song Lini, Zhang Yichen, Li Qi, Yang Jinkui. Establishment of MrgD knockout mice model with CRISPR/Cas9 gene targeting technology[J]. Journal of Capital Medical University, 2018, 39(4): 517-521.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2018.04.007

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2018/V39/I4/517

References

[1] 傅继梁. 基因工程小鼠(精)[M]. 上海:上海科技出版社, 2006.
[2] Li W, Teng F, Li T, et al. Simultaneous generation and germline transmission of multiple gene mutations in rat using CRISPR-Cas systems[J]. Nat Biotechnol, 2013, 31(8):684-686.
[3] Hsu P D, Lander E S, Zhang F. Development and applications of CRISPR-Cas9 for genome engineering[J]. Cell, 2014, 157(6):1262-1278.
[4] Platt R J, Chen S, Zhou Y, et al. CRISPR-Cas9 knockin mice for genome editing and cancer modeling[J]. Cell, 2014, 159(2):440-455.
[5] Burgess D J. Technology:a CRISPR genome-editing tool[J]. Nat Rev Genet, 2013, 14(2):80.
[6] Dong X, Han S, Zylka M J, et al. A diverse family of GPCRs expressed in specific subsets of nociceptive sensory neurons[J]. Cell, 2001, 106(5):619-632.
[7] Lembo P M, Grazzini E, Groblewski T, et al. Proenkephalin a gene products activate a new family of sensory neuron——specific GPCRs[J]. Nat Neurosci, 2002, 5(3):201-209.
[8] Shinohara T, Harada M, Ogi K, et al. Identification of a G protein-coupled receptor specifically responsive to beta-alanine[J]. J Biol Chem, 2004, 279(22):23559-23564.
[9] Santos R A, Simoes e silva A C, Maric C, et al. Angiotensin-(1-7) is an endogenous ligand for the G protein-coupled receptor Mas[J]. Proc Nat Acad Sci U S A, 2003, 100(14):8258-8263.
[10] Gembardt F, Grajewski S, Vahl M, et al. Angiotensin metabolites can stimulate receptors of the Mas-related genes family[J]. Mol Cell Biochem, 2008, 319(1-2):115-123.
[11] 康怡,张宝和,徐洪涛,等.北京市部分中老年男性高胰岛素血症患者代谢紊乱及胰岛功能评估[J].首都医科大学学报,2016,37(3):385-390.
[12] 史婷婷, 杨芳远, 曹曦,等. 血管紧张素转化酶2减少胰岛细胞的氧化应激及凋亡[J]. 首都医科大学学报, 2017, 38(2):151-155.
[13] Cao X, Yang F, Shi T, et al. Angiotensin-converting enzyme 2/angiotensin-(1-7)/Mas axis activates Akt signaling to ameliorate hepatic steatosis[J]. Sci Rep, 2016, 6:21592.
[14] 寻英, 秦映芬, 徐梦婕,等. ACE2-Ang-(1-7)-Mas轴对糖代谢作用的研究进展[J]. 广东医学, 2015(1):150-153.
[15] Hrenak J, Paulis L, Simko F. Angiotensin A/alamandine/MrgD axis:another clue to understanding cardiovascular pathophysiology[J]. Int J Mol Sci, 2016, 17(7):pii:E1098.
[16] Nishimura S, Uno M, Kaneta Y, et al. MRGD, a MAS-related G-protein coupled receptor, promotes tumorigenisis and is highly expressed in lung cancer[J]. PLoS One, 2012, 7(6):e38618.
[17] Crozier R A, Ajit S K, Kaftan E J, et al. MrgD activation inhibits KCNQ/M-currents and contributes to enhanced neuronal excitability[J]. J Neurosci, 2007, 27(16):4492-4496.

Establishment of MrgD knockout mice model with CRISPR/Cas9 gene targeting technology

PDF

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 8

Recommended Articles

Metrics

Comments

[1]	Chen Jianhong, Lin Xin, Feng Jinqiu, Ding Pengpeng, Wang Miaomiao, Lin Lin, Liu Hong, Wu Jing. Phenotypic analysis of hepatocyte-specific Eva1a/Tmem166 gene knockout mice [J]. Journal of Capital Medical University, 2021, 42(4): 568-574.
[2]	Xiong Fengran, Lu Jing, Zhang Yingchao, Xie Rongrong, Zhao Ruxuan, Li Qi, Yang Jinkui. Establishment of Kcnh6 gene site-specific mutant mice modeled on a pedigree with monogenic diabetes and the phenotype analysis [J]. Journal of Capital Medical University, 2021, 42(4): 575-581.
[3]	Gu Siyu, Yang Xiaomei, He Junqi. gene editing technology: gene scissors—a tool for rewriting the code of life—introduction of The Nobel Prize in Chemistry 2020 [J]. Journal of Capital Medical University, 2020, 41(6): 1014-1018.
[4]	Liu Xin, Xu Youqing, Cui Chunying, Zhao Zhigang. Application of GPIHBP1 gene knockout mice in the study of lung injury induced by acute pancreatitis with severe hypertriglyceridemia [J]. Journal of Capital Medical University, 2020, 41(4): 564-569.
[5]	Li Qi, Lu Jing, Zhu Xiaorong, Xiong Fengran, Yang Jinkui. Generation and identification of Unc13 gene pancreatic beta cell knockout mice model [J]. Journal of Capital Medical University, 2020, 41(1): 14-20.
[6]	Lu Jing, Wu Qian, Xie Rongrong, Qiu Haiyan, Cheng Cheng, Yang Jinkui. Construction and preliminary application of BACE2 gene knockout in zebrafish using CRISPR/Cas9 [J]. Journal of Capital Medical University, 2015, 36(6): 839-843.
[7]	Cheng Cheng, Wu Qian, Lu Jing, Qiu Haiyan, Li Qian, Yang Jinkui. Effect of RFX6 on pancreas development in zebrafish using CRISPR/Cas9 [J]. Journal of Capital Medical University, 2015, 36(6): 844-847.
[8]	DING Lei;GAO Hong;ZHU Yu-bing;LI Wen-bin. Claudin-7—a potential tumor suppressor gene [J]. Journal of Capital Medical University, 2012, 33(5): 569-574.