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Targeted mutagenesis of the murine IRP1 and IRP2 genes reveals context-dependent RNA processing differences in vivo.

We report the targeted mutagenesis of the murine iron regulatory protein (IRP)-1 and IRP2 genes, respectively, with a classical gene trap construct. Insertion of the targeting cassette into the second intron of either gene by homologous recombination interrupts their open reading frames near the N termini. Mice that are homozygous for the correctly modified IRP1 or IRP2 alleles, respectively, display a strong reduction (90%, IRP1(-/-)) or nondetectable levels (IRP2(-/-)) of the targeted proteins. Interestingly, the pre-mRNAs transcribed from the identical targeting cassettes are processed differently within the two different contexts. Detailed analysis of the respective products identifies the choice of alternative splice and 3' end processing sites in the same tissues in vivo. We discuss the implications for the understanding of RNA processing and for targeting strategies for functional genomics in the mouse.

Pubmed ID: 15208438


  • Galy B
  • Ferring D
  • Benesova M
  • Benes V
  • Hentze MW


RNA (New York, N.Y.)

Publication Data

July 21, 2004

Associated Grants


Mesh Terms

  • Alternative Splicing
  • Animals
  • Base Sequence
  • DNA Primers
  • Exons
  • Genome
  • Iron Regulatory Protein 1
  • Iron Regulatory Protein 2
  • Mice
  • Mutagenesis, Site-Directed
  • RNA
  • RNA Precursors
  • RNA Processing, Post-Transcriptional
  • RNA, Messenger
  • Recombination, Genetic
  • Restriction Mapping
  • Reverse Transcriptase Polymerase Chain Reaction