title: Sus scrofa matrix attachment region enhances expression of the PiggyBac system transfected into HEK293T cells.
abstract: OBJECTIVE
  To determine the effects of the Sus scrofa matrix attachment region (SusMAR) on transgene expression in HEK293T cells.
  
  
  RESULTS
  Three expression vectors with the MAR at different sites in the PiggyBac (PB) transposon vector backbone were compared: two MARs flanking the β-galactosidase (β-gal) expression cassette, and one at the upstream or downstream site. Bos taurus MAR (BosMAR) and a β-gal expression cassette without MARs were the positive and negative controls, respectively. Compared to the control, β-gal activity of all SusMAR and BosMAR vectors was significantly improved in the presence of PB transposase (PBase). However, only the downstream SusMAR and upstream BosMAR vectors showed increased expression in the absence of PBase. Expression was significantly increased in all vectors with the PBase group compared to those without the PBase group. Gene copy numbers were not increased compared to the negative control.
  
  
  CONCLUSIONS
  SusMAR enhanced recombinant gene expression levels and stability in HEK293T cells, was not increase transgene copy number. These results could facilitate the development of vectors for stable production of therapeutic proteins.
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China.
authors.forename: Xin-Jian
authors.initials: XJ
authors.lastname: Wang
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China.
authors.forename: Jiang
authors.initials: J
authors.lastname: Wang
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China.
authors.forename: Yue-Ying
authors.initials: YY
authors.lastname: Wang
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China.
authors.forename: Yu-Jie
authors.initials: YJ
authors.lastname: Guo
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China.
authors.forename: Bei-Bei
authors.initials: BB
authors.lastname: Chu
authors.affiliation: College of Animal Husbandary and Veterinary Science, Henan Agricultural University, Wenhua Road 95, Zhengzhou, 450002, People's Republic of China. 516689530@qq.com.
authors.forename: Guo-Yu
authors.initials: GY
authors.lastname: Yang
mesh_terms: D000818:Animals; D004251:DNA Transposable Elements; D018628:Gene Dosage; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D045170:Matrix Attachment Regions / Q000235:genetics*; D010957:Plasmids; D034421:Sus scrofa / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D019076:Transgenes; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D004251:DNA Transposable Elements; D001616:beta-Galactosidase
keywords: Matrix attachment region; PiggyBac transposon; Therapeutic protein production; Transgene copy number; Transgene expression