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Proximity-dependent and proximity-independent trans-splicing in mammalian cells.

Publication ,  Journal Article
Viles, KD; Sullenger, BA
Published in: RNA
June 2008

Most human pre-mRNAs are cis-spliced, removing introns and joining flanking exons of the same RNA molecule. However, splicing of exons present on separate pre-mRNA molecules can also occur. This trans-splicing reaction can be exploited by pre-trans-splicing molecules (PTMs), which are incapable of cis-splicing. PTM-mediated trans-splicing has been utilized to repair mutant RNAs as a novel approach to gene therapy. Herein we explore how the site of PTM expression influences trans-splicing activity. We stably inserted a PTM expression cassette into the genome of HEK293 cells, generating clonal lines with single, unique insertion sites. We analyzed trans-splicing to the gene where the PTM was integrated, as well as genes neighboring these loci. We observed some pre-mRNAs only serve as substrates for trans-splicing when they are expressed in immediate proximity to the PTM expression site. The need for PTMs to be in close proximity with pre-mRNAs to trans-splice with them is consistent with the observation that pre-mRNA cis-splicing occurs cotranscriptionally. Interestingly, we identified several cellular pre-mRNAs in one localized area that serve as trans-splicing substrates irrespective of the PTM expression site. Thus, we find multiple cellular pre-mRNAs require PTM expression in close proximity to trans-splice while others do not.

Duke Scholars

Published In

RNA

DOI

EISSN

1469-9001

Publication Date

June 2008

Volume

14

Issue

6

Start / End Page

1081 / 1094

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Trans-Splicing
  • Spliceosomes
  • Rats
  • RNA Precursors
  • Humans
  • Exons
  • Developmental Biology
  • Cell Line
  • Animals
 

Citation

APA
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ICMJE
MLA
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Viles, K. D., & Sullenger, B. A. (2008). Proximity-dependent and proximity-independent trans-splicing in mammalian cells. RNA, 14(6), 1081–1094. https://doi.org/10.1261/rna.384808
Viles, Kristi D., and Bruce A. Sullenger. “Proximity-dependent and proximity-independent trans-splicing in mammalian cells.RNA 14, no. 6 (June 2008): 1081–94. https://doi.org/10.1261/rna.384808.
Viles KD, Sullenger BA. Proximity-dependent and proximity-independent trans-splicing in mammalian cells. RNA. 2008 Jun;14(6):1081–94.
Viles, Kristi D., and Bruce A. Sullenger. “Proximity-dependent and proximity-independent trans-splicing in mammalian cells.RNA, vol. 14, no. 6, June 2008, pp. 1081–94. Pubmed, doi:10.1261/rna.384808.
Viles KD, Sullenger BA. Proximity-dependent and proximity-independent trans-splicing in mammalian cells. RNA. 2008 Jun;14(6):1081–1094.

Published In

RNA

DOI

EISSN

1469-9001

Publication Date

June 2008

Volume

14

Issue

6

Start / End Page

1081 / 1094

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Trans-Splicing
  • Spliceosomes
  • Rats
  • RNA Precursors
  • Humans
  • Exons
  • Developmental Biology
  • Cell Line
  • Animals