The 3'UTR of HIC mRNA improves the production of recombinant proteins in Chinese hamster ovary cells.
Positive transcription elongation factor b (P-TEFb) is an important transcriptional regulator which controls 70-80% of RNA polymerase II transcription. It has been reported that the human I-mfa (inhibitor of MyoD family a) domain-containing protein (HIC) interacts with P-TEFb and that expression of HIC cDNA stimulates P-TEFb-dependent transcription. Interestingly, our recent study shows that transcriptional stimulation by HIC is predominately due to the 3' untranslated region (3'UTR) of HIC mRNA rather than its coding region. In this report, we investigate the effects of HIC 3'UTR on recombinant protein expression in mammalian cells. In transient transfections, overexpression of HIC 3'UTR stimulates transgene expression in several mammalian cell lines and significantly increases the production of human erythropoietin and interferon-gamma in Chinese hamster ovary (CHO) cells. This is the first report that demonstrates the improvement of expression of biopharmaceutical proteins by overexpressing a non-coding 3'UTR in CHO cells.
Liu
Jaron
J
Expression Engineering Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore,
Ku
Sebastian C Y
SC
Lee
Jialing
J
Young
Tara M
TM
Pe'ery
Tsafi
T
Mathews
Michael B
MB
Chao
Sheng-Hao
SH
D020413:3' Untranslated Regions; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004921:Erythropoietin / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000235:genetics; D008954:Models, Biological; D018006:Myogenic Regulatory Factors / Q000235:genetics* / Q000378:metabolism; D042925:Positive Transcriptional Elongation Factor B / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D019076:Transgenes
D020413:3' Untranslated Regions; C478034:MDFIC protein, human; D018006:Myogenic Regulatory Factors; D012333:RNA, Messenger; D011994:Recombinant Proteins; D004921:Erythropoietin; D007371:Interferon-gamma; D042925:Positive Transcriptional Elongation Factor B