Porcine Type III RNA Polymerase III Promoters for Short Hairpin RNA Expression

作者：Chuang Chin kai^*; Lee Kun Hsiung; Fan Chiu Tin; Su Yu Show

来源：Animal Biotechnology, 2009, 20(1): 34-39.

DOI：10.1080/10495390802603064

• shRNA
• Swine
• Type III RNA Polymerase III Promoter

Based on the highly conserved sequences of small nuclear RNA and small cytoplasmic RNA between vertebrate species, three porcine type III RNA polymerase III promoters, pY1, pY3 and pU6, were identified by using genomic DNA walking. To test the functional relationship of these sequences, the human H1 promoter of pSUPER-EGFP-l-neo vector was substituted with these three promoters to create the ppPol III-MCS vectors. The strength of each promoter was measured by its ability to derive expression of shRNA to repress expression of luciferase via RNA interference in the pig kidney epithelial cell line LLC-PK1. We determine that the ranking of promoter strength in descending order is pU6pY1pY3.

• 出版日期2009