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. 1989 Nov 3;59(3):531-9.
doi: 10.1016/0092-8674(89)90036-6.

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

B J Blencowe  1 , B S Sproat, U Ryder, S Barabino, A I Lamond
Affiliations

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

B J Blencowe et al. Cell. .

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.

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