Establishment of Hela229 Stable Cell Lines with Interleukin-1 Alpha Gene Knockdown

Objective: Constructing IL-1α gene targeted short hairpin RNA interference expression vector,which can inhibit the endogenous expression of IL-1α of Hela229 cells,to establish the endogenous IL-1α gene knockdown stable cell line. Method: Design a 21 bp target nucleotide sequence according to the principle of shRNA desigation,clone the DNA template of siRNA into the shRNA expression vector pRNAT U6. 1 / Neo,gain the inhibition of IL-1α gene recombinant shRNA expression plasmid,transfect the plasmid into Hela229 cells. Monoclonal stable cell lines was established after screening by G418. Detect the IL-1α supression effect at the protein level via the method of ELISA. Results: IL-1α-shRNA carrier recombination were determined the correct construction through the enzyme identification and sequencing analysis. ELISA results showed that the restructuring of the IL-1α-shRNA expression vector can significantly inhibit the expression of endogenous IL-1α of Hela229 cells. Conclusion: Targeted IL-1α gene recombinant shRNA expression plasmid can significantly inhibit the expression of endogenous IL-1α of Hela229 cells. Successfully established the Hela229 stable cell line with IL-1α gene knockdown.