Assaying nuclear messenger RNA export in human cells.
Journal Article (Journal Article)
This chapter describes a simple method for the analysis of nuclear messenger RNA (mRNA) export in human cells in culture. The assay described relies on the observation that mRNA molecules containing an intron are generally retained in the nucleus until splicing is completed. Upon sequestration of the cat indicator gene in a single intron located 5' to an mRNA cap site, CAT protein expression becomes dependent on the specific recruitment of a nuclear RNA export factor to the unspliced cat RNA via an inserted RNA binding site. This site can be a natural, high affinity RNA target for the nuclear export factor or alternately the export factor can be tethered to the unspliced cat mRNA by fusion to a heterologous RNA binding domain.
Full Text
Cited Authors
- Cullen, BR
Published Date
- 2004
Published In
Volume / Issue
- 257 /
Start / End Page
- 85 - 92
PubMed ID
- 14769998
International Standard Serial Number (ISSN)
- 1064-3745
Digital Object Identifier (DOI)
- 10.1385/1-59259-750-5:085
Language
- eng
Conference Location
- United States