Modification of the pig CFTR gene mediated by small fragment homologous replacement

Ann N Y Acad Sci. 2006 Oct:1082:120-3. doi: 10.1196/annals.1348.063.

Abstract

The generation of a pig model of cystic fibrosis (CF) is a multistep process. Initial steps in this process involved the design and cloning of a small DNA fragment (SDF) or large oligodeoxynucleotide (LODN) that contains the F508del mutation and a silent restriction fragment length polymorphism causing mutation. This SDF/LODN was transfected into wild-type (WT) pig fetal fibroblast with the intention of modifying the pig genomic DNA by small fragment homologous replacement (SFHR). The targeted deletion (F508del) was detected in a subpopulation of transfected cells by allele-specific polymerase chain reaction (AS-PCR).

MeSH terms

  • Animals
  • Cells, Cultured
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
  • Cystic Fibrosis*
  • Disease Models, Animal*
  • Fibroblasts / metabolism
  • Gene Targeting / methods*
  • Oligodeoxyribonucleotides / genetics
  • Polymerase Chain Reaction
  • Polymorphism, Restriction Fragment Length
  • Sequence Deletion
  • Swine
  • Transfection

Substances

  • Oligodeoxyribonucleotides
  • Cystic Fibrosis Transmembrane Conductance Regulator