Lampe1: An ENU-germline mutation causing spontaneous hepatosteatosis identified through targeted exon-enrichment and next-generation sequencing

Rachel Sheridan, Kristin Lampe, Shiva Kumar Shanmukhappa, Patrick Putnam, Mehdi Keddache, Senad Divanovic, Jorge Bezerra, Kasper Hoebe

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

Using a small scale ENU mutagenesis approach we identified a recessive germline mutant, designated Lampe1 that exhibited growth retardation and spontaneous hepatosteatosis. Low resolution mapping based on 20 intercrossed Lampe1 mice revealed linkage to a ~14 Mb interval on the distal site of chromosome 11 containing a total of 285 genes. Exons and 50 bp flanking sequences within the critical region were enriched with sequence capture microarrays and subsequently analyzed by next-generation sequencing. Using this approach 98.1 percent of the targeted DNA was covered with a depth of 10 or more reads per nucleotide and 3 homozygote mutations were identified. Two mutations represented intronic nucleotide changes whereas one mutation affected a splice donor site in intron 11-12 of Palmitoyl Acetyl-coenzyme A oxygenase-1 (Acox1), causing skipping of exon 12. Phenotyping of Acox1Lampe1 mutants revealed a progression from hepatosteatosis to steatohepatitis, and ultimately hepatocellular carcinoma. The current approach provides a highly efficient and affordable method to identify causative mutations induced by ENU mutagenesis and animal models relevant to human pathology.

Original languageEnglish (US)
Article numbere21979
JournalPloS one
Volume6
Issue number7
DOIs
StatePublished - 2011
Externally publishedYes

ASJC Scopus subject areas

  • General

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