Sequence-specific affinity selection of mammalian splicing complexes

Ursula Ryder, Brian S. Sproat, Angus I. Lamond (Lead / Corresponding author)

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)

Abstract

Antisense oligonucleotides made of 2′-OMe RNA are shown to bind specifically and efficiently to targeted sites on pre-mRNA substrates, allowing affinity selection of splicing complexes using streptavidin/blotin chromatography. The position of probe binding to the pre-mRNA influences which type of splicing complex can be selected. The accessibility of pre-mRNA sequences to antisense probes changes during the course of the splicing reaction. U1, U2, U4, U5 and U6 snRNAs are all detected in affinity-selected mammalian splicing complexes. However, antisense oligonucleotides targeted to snRNAs can block the binding of specific snRNPs to pre-mRNA. Quantitative affinity selection analyses show that only a small fraction of snRNPs in a HeLa nuclear splicing extract participate in spliceosome formation.

Original languageEnglish
Pages (from-to)7373-7379
Number of pages7
JournalNucleic Acids Research
Volume18
Issue number24
DOIs
Publication statusPublished - 1 Dec 1990

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