Sequence-specific affinity selection of mammalian splicing complexes.

U Ryder; B S Sproat; A I Lamond

doi:10.1093/nar/18.24.7373

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PubMed · 2147994

Sequence-specific affinity selection of mammalian splicing complexes.

U Ryder·B S Sproat·A I Lamond

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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/biotin 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.

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Base Sequence Chromatography, Affinity HeLa Cells Humans Molecular Sequence Data Oligoribonucleotides RNA Precursors RNA Splicing RNA, Antisense RNA, Small Nuclear Ribonucleoproteins Ribonucleoproteins, Small Nuclear Journal Article

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U Ryder, B S Sproat, A I Lamond. 1990-12-25. Sequence-specific affinity selection of mammalian splicing complexes.. https://doi.org/10.1093/nar%2F18.24.7373

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