When good transcripts go bad: artifactual RT-PCR 'splicing' and genome analysis

Bioessays. 2008 Jun;30(6):601-5. doi: 10.1002/bies.20749.

Abstract

Gene and intron prediction are essential for accurate inferences about genome evolution. Recently, two genome-wide studies searched for recent intron gains in humans, reaching very different conclusions: either of a complete absence of intron gain since early mammalian evolution, or of creation of numerous introns by genomic duplication in repetitive regions. We discuss one possible explanation: the underappreciated phenomenon of "template switching", by which reverse transcriptase may create artifactual splicing-like events in the preparation of cDNA/EST libraries, may cause complications in searches for newly gained introns in repetitive regions. We report large numbers of apparent template switching in transcript sequences from the intron-poor protists Trichomonas vaginalis and Giardia lamblia. Supplementary material for this article can be found on the BioEssays website (http://www.interscience.wiley.com/jpages/0265-9247/suppmat/index.html).

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Alleles
  • Animals
  • Base Sequence
  • DNA, Protozoan / genetics
  • Evolution, Molecular*
  • Gene Duplication
  • Genomics
  • Giardia lamblia / genetics
  • Giardia lamblia / metabolism
  • Humans
  • Introns*
  • Models, Genetic
  • Molecular Sequence Data
  • RNA Splicing / genetics*
  • RNA-Directed DNA Polymerase / metabolism
  • Repetitive Sequences, Nucleic Acid
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription, Genetic
  • Trichomonas vaginalis / genetics
  • Trichomonas vaginalis / metabolism

Substances

  • DNA, Protozoan
  • RNA-Directed DNA Polymerase