Dec 25, 1990

Sequence-specific affinity selection of mammalian splicing complexes

Nucleic Acids Research
U RyderA I Lamond

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.

Mentioned in this Paper

Small Nuclear RNA
Primary RNA Transcript
Complex (molecular entity)
Chromatography
Antisense RNA
Small Nuclear Ribonucleoproteins
Antisense Probes
Heterogeneous Nuclear RNA
Biotin
Nuclear mRNA Cis Splicing, via Spliceosome

About this Paper

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