Enhanced expression of transgene in CHO cells using matrix attachment region.
The expression of transgenes in mammalian cells is often at a low level mainly due to position effects from the neighboring chromatin context. To improve this, we have constructed a vector pCAM, which contains chloramphenicol acetyltransferase (CAT) reporter gene cassettes, driven by SV40 early promoter and flanked by two human beta-globin MARs in cis. We transfected this vector into the Chinese hamster ovary (CHO) cell line, and found that the level of CAT gene expression with MAR was effectively increased, about 5.493-fold higher than those without MARs. Moreover, the variations of CAT expression among individuals of transformants were decreased 2.670-fold. Our result also showed that MAR could increase the proportion of positive colonies in recombinants.
Wang
Tian-Yun
TY
Department of Biochemistry and Molecular Biology, Jinsui Road, Xinxiang, Henan 453003, PR China. wty@xxmu.edu.cn
Yang
Rui
R
Qin
Chuan
C
Wang
Li
L
Yang
Xian-Jun
XJ
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D018628:Gene Dosage; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D014162:Transfection; D019076:Transgenes