Nov 3, 1989

Antisense probing of the human U4/U6 snRNP with biotinylated 2'-OMe RNA oligonucleotides

Cell
B J BlencoweA I Lamond

Abstract

We have used antisense 2'-OMe RNA oligonucleotides carrying four 5'-terminal biotin residues to probe the structure and function of the human U4/U6 snRNP. Nine oligonucleotides, complementary to multiple regions of U4 and U6 snRNAs, bound stably and specifically to U4/U6 snRNP. This allowed for efficient and selective removal of U4/U6 from HeLa cell nuclear extracts. Binding of oligonucleotides to certain snRNA domains inhibited splicing and affected the U4-U6 interaction. Pre-mRNA and splicing products could also be affinity-selected through binding of the oligonucleotides to U4/U6 snRNPs in splicing complexes. The results suggest that U4 snRNP is not released during spliceosome assembly.

Mentioned in this Paper

RNA Conformation
Biodermatin
Antisense RNA
Small Nuclear Ribonucleoproteins
Methylation
Oligoribonucleotide Probes
Northern Blot
Genomic Hybridization
Poly(A) Tail
RNA, Messenger, Splicing

About this Paper

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