A yeast-based recombinogenic targeting toolset for transgenic analysis of human disease genes

Kevin L. Bentley, Cooduvalli S. Shashikant, Wayne Wang, Nancy H. Ruddle, Frank H. Ruddle

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Transgenic mousemodels are valuable resources for analyzing functions of genes involved in human diseases. Mouse models provide critical insights into biological processes, including in vivo visualization of vasculature critical to our understanding of the immune system. Generating transgenic mice requires the capture and modification of largeinsert DNAs representing genes of interest. We have developed a methodology using a yeast-bacterial shuttle vector, pClasper, that enables the capture and modification of bacterial artificial chromosomes (BAC)-sized DNA inserts. Numerous improvements and technical advances in the original pClasper vector have allowed greater flexibility and utility in this system. Examples of such pClasper mediated gene modifications include: Claspette-mediated capture of large-insert genomic fragments from BACs-human polycystic kidney disease-1 (PKD1); modification of pClasperA clones by the RareGap method-PKD1 mutations; Claspette-mediated modification of pClasper clones- mouse albumin-1 gene; and, of most relevance to our interest in lymph node vasculature-Claspimer-mediated modification of pClasper clones-high endothelial venule and lymphatic vessel genes. Mice that have been generated with these methods include mice with fluorescent high endothelial venules.

Original languageEnglish (US)
Pages (from-to)E58-E68
JournalAnnals of the New York Academy of Sciences
Volume1207
Issue numberSUPPL.1
DOIs
StatePublished - Jan 1 2010

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • History and Philosophy of Science

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