Porcine type III RNA polymerase III promoters for short hairpin RNA expression.
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 pU6 > pY1 > pY3.
Chuang
Chin-kai
CK
Division of Biotechnology, Animal Technology Institute Taiwan, Miaoli, Taiwan. jkjuang@mail.atit.org.tw
Lee
Kun-Hsiung
KH
Fan
Chiu-Tin
CT
Su
Yu-Show
YS
D000818:Animals; D001483:Base Sequence; D005786:Gene Expression Regulation; D055029:Inverted Repeat Sequences; D008967:Molecular Biology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D012313:RNA / Q000235:genetics*; D012320:RNA Polymerase III / Q000235:genetics*; D013552:Swine / Q000235:genetics*
D012313:RNA; D012320:RNA Polymerase III