title: Functional analysis of SH3 domain containing ring finger 2 during the myogenic differentiation of quail myoblast cells.
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 vivoandin vitrofunctional genomic analyses should be a prerequisite.
  
  
  METHODS
  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,SH3RF2was knocked out using clustered regularly interspaced short palindromic repeat/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.
  
  
  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 inSH3RF2. 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea.
authors.forename: Si Won
authors.initials: SW
authors.lastname: Kim
authors.affiliation: Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea.
authors.forename: Jeong Hyo
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea.
authors.forename: Tae Sub
authors.initials: TS
authors.lastname: Park
keywords: Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-Cas9; Functional Genomics; Knockout; Muscle Differentiation; Myoblast; SH3 Domain Containing Ring Finger 2 (SH3RF2)