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Asian-Australas J Anim Sci > Accepted
doi: https://doi.org/10.5713/ajas.16.0865    [Accepted]
Functional analysis of SH3 domain containing ring finger 2 (SH3RF2) during the myogenic differentiation of QM7 cells
Si Won Kim, Jeong Hyo Lee, Tae Sub Park*
Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang-gun, Gangwon-do 25354, Korea
* Corresponding Author: Tae Sub Park ,Tel: 033-339-5721, Fax: 033-339-5763, Email: taesubpark@snu.ac.kr
Received: 11 November 2016;  Revised: 14 December 2016.  Accepted: 21 December 2016.  Published online 27 December 2016.

ABSTRACT
Objective:
Owing to the public availability of complete genome sequences, including avian species, massive bioinformatics analyses may be conducted for computational gene prediction and the identification of gene regulatory networks through various informatics tools. However, to evaluate the biofunctional activity of a predicted target gene, in vivo and in vitro functional genomic analyses should be a prerequisite.
Method:
Due to a lack of quail genomic sequence information, we first identified the partial genomic structure and sequences of the quail SH3 domain containing ring finger 2 (SH3RF2) gene. Subsequently, SH3RF2 was knocked out using CRISPR/Cas9 technology and single cell-derived SH3RF2 mutant sublines were established to study the biofunctional activity of SH3RF2 in quail myoblast (QM7) cells during muscle differentiation. Through a T7 endonuclease I assay and genotyping analysis, we established an SH3RF2 knockout (KO) QM7#4 subline with 61 and 155 nucleotide deletion mutations in SH3RF2. After the induction of myotube differentiation, the expression profiles were analyzed and compared between regular QM7 and SH3RF2 KO QM7#4 cells by global RNA sequencing and bioinformatics analysis.
Conclusion:
We did not detect any statistically significant role of SH3RF2 during myotube differentiation in QM7 myoblast cells. However, additional experiments are necessary to examine the biofunctional activity of SH3RF2 in cell proliferation and muscle growth.
Keywords: CRISPR-Cas9; Knockout; Myoblast; Muscle differentiation; SH3RF2; Functional genomics
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Proteomic Analysis of Bovine Muscle Satellite Cells during Myogenic Differentiation  2011 September;24(9)
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