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Modification of the Pig CFTR Gene Mediated by Small Fragment Homologous Replacement
Author(s) -
MAURISSE ROSALIE,
CHEUNG JUDY,
WIDDICOMBE JONATHAN,
GRUENERT DIETER C.
Publication year - 2006
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1348.063
Subject(s) - microbiology and biotechnology , transfection , homologous recombination , genomic dna , biology , polymerase chain reaction , restriction fragment length polymorphism , mutant , dna , gene , chemistry , genetics
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)