title: [Knocking-out extra domain A alternative splice fragment of fibronectin using a clustered regularly interspaced short palindromic repeats/associated proteins 9 system].
abstract: OBJECTIVE
  To investigate the effect of the fibronectin extra domain A on the aggressiveness of salivary adenoid cystic carcinoma (SACC) cells, via the clustered regularly interspaced short palindromic repeats (CRISPR)/ associated proteins (Cas) system.
  
  
  METHODS
  One sgRNA was designed to target the upstream of the genome sequences of extra domain A(EDA) exon and the downstream. Then the sgRNA was linked into plasmid PX-330 and transfected into SACC-83 cells. PCR and DNA sequence were used to testify the knockout cells, and the monoclones of EDA absent SACC cells were selected (A+C-2, A+C-6, B+C-10). CCK-8 cell proliferation and invasion was then tested in control group and the experimental group.
  
  
  RESULTS
  The sgRNA was successfully linked into PX-330 plasmid. Part of adenoid cystic carcinoma cells' SACC-83 genomic EDA exon was knocked out, and the knockdown efficiency was above 70%, but the total amount of fibronectin did not change significantly. Three monoclones of EDA absent SACC- 83 cells were successfully selected with diminished migration and proliferation.
  
  
  CONCLUSIONS
  The CRISPR/Cas9 system was a simplified system with relatively high knockout efficiency and EDA knockout could inhibiting SACC cell's mobility and invasiveness.
authors.affiliation: The Central Laboratory, Peking University School and Hospital of Stomatology, Beijing 100081, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Yang
authors.forename: Haicheng
authors.initials: H
authors.lastname: Wang
authors.forename: Shuyu
authors.initials: S
authors.lastname: Xu
authors.forename: Jing
authors.initials: J
authors.lastname: Peng
authors.forename: Jiuhui
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Email: licuiying_67@163.com.
authors.forename: Cuiying
authors.initials: C
authors.lastname: Li
mesh_terms: D001483:Base Sequence; D003528:Carcinoma, Adenoid Cystic / Q000235:genetics* / Q000473:pathology*; D002465:Cell Movement / Q000235:genetics; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D005091:Exons; D005353:Fibronectins / Q000737:chemistry / Q000235:genetics*; D055786:Gene Knockout Techniques; D006801:Humans; D009361:Neoplasm Invasiveness; D010957:Plasmids / Q000235:genetics*; D012313:RNA; D014162:Transfection
chemical_list: D005353:Fibronectins; D012313:RNA