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Bioinformatic Identification of Chinese Hamster Ovary (CHO) Cold-Shock Genes and Biological Evidence of their Cold-Inducible Promoters.
Biotechnology Journal ( IF 4.7 ) Pub Date : 2019-12-11 , DOI: 10.1002/biot.201900359
Ly Ngoc Nguyen 1, 2 , Neža Novak 1, 2 , Martina Baumann 1 , Jadranka Koehn 3 , Nicole Borth 2
Affiliation  

Lower cultivation temperature dramatically affects cell growth and cellular productivity in Chinese hamster ovary cells (CHO) and is often used in industrial applications with the aim to enhance productivity. Cold-inducible proteins whose activity is induced at lower temperature play an important role in understanding the mechanisms of cold-induced changes in gene expression. One of these mechanisms is increased transcription of specific target genes controlled by sequence elements in cold-inducible promoters. This study provides a list of cold-inducible genes and endogenous cold-inducible promoters of CHO cells. Transcriptome data before and after a temperature shift from 37 to 33 °C are analyzed to identify 94 cold-inducible genes, which are highly expressed and have a high fold change in expression after the temperature shift. Cold-inducible promoters are identified from the top ten cold-inducible genes, showing up to 11-fold increased luciferase expression at lowered temperature in transient transfections. Additionally, several common transcription factor binding sites are identified in the top cold-inducible promoter sequences and expression of their corresponding transcription factors over temperature-shift cultivation is evaluated. These may be responsible for enhanced promoter activity under lower temperature and can be used as engineering targets.
更新日期:2019-12-11
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