Skip to main content

Advertisement

Table 1 Analysis of differential gene expression in HeLa triggered by different cultivation temperatures.

From: Novel approaches to render stable producer cell lines viable for the commercial manufacturing of rAAV-based gene therapy vectors

Name Differential expression at Mode of regulation Microarray analysis RT qPCR
Gene A 30°C Up 3.2-fold 6.9-fold
Gene B 30°C Up 2.2-fold 2.6-fold
Gene C 30°C Up 3.3-fold 2.3-fold
miRNA A 32°C Up 3.1-fold -
miRNA B 32°C Down 3.3-fold -
miRNA C 32°C Up 3.0-fold -
  1. Cells were seeded at two different densities and cultivated at 37°C for two days. Subsequently, cells were incubated for another 6 hours at 30, 32, and 37°C, respectively, before mRNA was isolated from the cells. Microarray analysis (GeneChip® Human Exon 1.0 ST Array, Affymetrics) was performed to identify mRNAs differentially expressed more than 2-fold. Validation was done by RT qPCR analysis (EvaGreen® Mastermix, Biorad) and included controls of regulated and non-regulated mRNAs [12, 11, 1]. Differentially expressed miRNAs (>2-fold) were also identified by microarray analysis (GeneChip® miRNA 2.0 Array, Affymetrics). As validation is not yet completed, only an excerpt of the most promising miRNA candidates is shown.