CRISPR/Cas9介导RB1基因敲除及其在鸡前脂肪细胞分化、增殖中的功能研究

doi:10.11843/j.issn.0366-6964.2016.09.005

Abstract

Abstract:

In order to study the role of RB1 gene on chicken preadipocyte differentiation and proliferation.CRISPR/Cas9 system was established and conducted to knockout the RB1 gene in the chicken preadipocyte.The proliferation and differentiation of chicken preadipocytes and the expression of related marker genes were detected.Results showed that the CRISPR/Cas9 system could effectively mediate RB1 knockout and cause truncated translation in chicken at the rate of 10%.Oil Red O and CCK-8 were used to evaluate the ability of differentiation and proliferation in chicken preadipocyte respectively.Knockout of RB1 in chicken preadipocyte resulted in enhanced proliferation capability and decreased differentiation capability in chicken preadipocyte.RB1 knockout decreased the transcriptional expression of adipocyte differentiation associated genes such as G0S2，FAS and A-FABP.Especially，the expression of G0S2 was significantly decreased at all stages of preadipocyte differentiation.Further， RB1 knockout increased the expression of cell proliferation associated genes such as CyclinD1，E2F1，Ki67 and PCNA, which was corresponded to the enhanced proliferation capability in the RB1 knockout cells.In summary，RB1 may be a key regulator on differentiation and proliferation of chicken preadipocyte.

CLC Number:

S831.2

ZHANG Qi，HUANG Jiao-jiao，YANG Cai-xia，WANG Yu-xiang，ZHANG Hui，ZHAO Jian-guo，LI Hui. CRISPR/Cas9 Mediated RB1 Knockout and Its Impact on Chicken Preadipocytes Differentiation and Proliferation[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(9): 1775-1784.

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CRISPR/Cas9 Mediated RB1 Knockout and Its Impact on Chicken Preadipocytes Differentiation and Proliferation

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