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Asian-Australas J Anim Sci > Volume 30(8); 2017 > Article
Asian-Australasian Journal of Animal Sciences 2017;30(8): 1183-1189. doi: https://doi.org/10.5713/ajas.16.0865
Functional analysis of SH3 domain containing ring finger 2 during the myogenic differentiation of quail myoblast cells
Si Won Kim1, Jeong Hyo Lee1, Tae Sub Park1,*
1Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea
* Corresponding Author: Tae Sub Park ,Tel: +82-33-339-5721, Fax: +82-33-339-5763, Email: taesubpark@snu.ac.kr
Received: November 11, 2016;  Revised: December 14, 2016.  Accepted: December 21, 2016.  Published online December 27, 2016.
Si Won Kim and Jeong Hyo Lee contributed equally to this work.

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 clustered regularly interspaced short palindromic repeat/Cas9 technology and single cell­derived SH3RF2 mutant sublines were established to study the biofunctional activity of SH3RF2 in quail myoblast (QM7) cells during muscle differentiation.
Results:
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 differ entiation in QM7 myoblast cells. However, additional experiments are necessary to examine the biofunctional activity of SH3RF2 in cell proliferation and muscle growth.
Keywords: Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)­Cas9; Knockout; Myoblast; Muscle Differentiation; SH3 Domain Containing Ring Finger 2 (SH3RF2); Functional Genomics
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