Mir 22-3p enhances recombinant protein production from HEK 293 cell by affecting the expression of HIPK1 and FRAT2

Improving protein production in human embryonic kidney cells is important structural studies and antibody production. The use of small non-protein coding RNA, such as microRNA, has been an effective method for increasing protein expression. Our high-throughput human microRNA screen in HEK 293 cells previously identified miRNA 22-3p as a promising candidate for increasing the expression of luciferase, couple of membrane proteins and a secreted fusion protein. To explore the mechanisms of this increase in protein expression and to understand the intracellular events, we conducted a gene expression analysis of luciferase- expressing HEK 293 cells transfected with a mir-22-3p mimic and compared with cells transfected with a negative control. Following microarray analysis, down-regulated genes were identified and were cross-referenced with the predicted targets of mir-22-3p and with results from our previous high-throughput siRNA screen. By performing common seed analysis on the possible targets, the list was narrowed to two genes, HIPK1 and FRAT2. These two genes were validated as being involved in improving luciferase production using siRNA and qRT PCR. Identification of genes expressed in WT vs.human embryonic kidney cells that were treated with Mir 22-3p by triplicated sample per group