Modified U1 snRNA suppresses expression of a targeted endogenous RNA by inhibiting polyadenylation of the transcript.
We have previously demonstrated that a modified U1 snRNA inhibits expression of a number of targeted transgenes. Here we exploit the ability of the modified U1 snRNA to inhibit endogenous gene expression and define the mechanism responsible for this inhibitory action. MC3T3-E1 cells stably transfected with U1 anti-Cbfa1 show a change of morphology from polygonal to fibroblast-like cells. This visual observation was supported by an 80% reduction of Cbfa1 expression and suppression of downstream genes associated with osteoblast differentiation. In rat ROS 17/2.8 cells, osteocalcin and Col1a1 gene expression was reduced up to 90% by the U1 anti-osteocalcin or U1 anti-Col1a1 construct, respectively. The length of mature osteocalcin mRNA poly(A) tail was significantly shortened in the targeted mRNA by transcript-specific poly(A) test (PAT)-PCR. These data demonstrate that the modified U1 snRNA is able to reduce endogenous gene expression by limiting the polyadenylation of the targeted pre-mRNA transcript.
Liu
Peng
P
Department of Genetics and Developmental Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA.
Kronenberg
Mark
M
Jiang
Xi
X
Rowe
David
D
D000818:Animals; D002460:Cell Line; D024042:Collagen Type I / Q000096:biosynthesis / Q000235:genetics; D000090602:Collagen Type I, alpha 1 Chain; D050659:Core Binding Factor Alpha 1 Subunit; D005786:Gene Expression Regulation; D005818:Genetic Engineering; D051379:Mice; D016296:Mutagenesis; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D015675:Osteocalcin / Q000096:biosynthesis / Q000235:genetics; D026723:Polyadenylation; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism*; D012342:RNA, Small Nuclear / Q000235:genetics*; D051381:Rats; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics
D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D050659:Core Binding Factor Alpha 1 Subunit; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D012342:RNA, Small Nuclear; D014157:Transcription Factors; C405483:U1 small nuclear RNA; D015675:Osteocalcin