ABCMdb

PMID: 14613541

Li Z, Engelhardt JF
Progress toward generating a ferret model of cystic fibrosis by somatic cell nuclear transfer.
Reprod Biol Endocrinol. 2003 Nov 7;1:83., 2003-11-07 [PubMed]

Sentences

No. Mutations Sentence Comment

106 ABCC7 p.Gly551Asp Since chimeraplasts are most efficient at introducing single base-pair alterations, we have evaluated the ability of chimeraplasts to target the G551D mutation to the CFTR gene that requires only a single-base alteration. Login to comment 107 ABCC7 p.Gly551Asp With the ultimate goal of generating ferret fibroblast cell lines heterozygous for the G551D mutation, we first sought to develop targeting and screening methodologies in Hela cells. Login to comment 123 ABCC7 p.Gly551Asp The sequences for the wild-type and mutant ASO primers were as follows: 1) wild-type, GAGT- GGAGGTCAACGAG, and 2) G551D mutant, GAGTGGA- GATCAACGAG. Login to comment 133 ABCC7 p.Gly551Asp ASO screening of 44 single-cell clones generated from chimeraplast-transfected Hela cells resulted in 3 positive clones that strongly hybridized to the G551D mutant oligonucleotide (Figure 4b and 4c). Login to comment 144 ABCC7 p.Gly551Asp ABCC7 p.Gly551Asp Recent reports have suggested that the polarity of the DNA segment in the Chimeraplast targeting of the G551D mutation to the humanCFTR gene in Hela cellsFigure 4 Chimeraplast targeting of the G551D mutation to the human CFTR gene in Hela cells. Login to comment 148 ABCC7 p.Gly551Asp ABCC7 p.Gly551Asp (B, C) Cell lysates prepared with 44 targeted Hela cell clones after G551D chimeric oligonucleotide transfection were amplified by 2 rounds of PCR and analyzed by ASO hybridization against (B) wild-type CFTR and (C) G551D mutant oligonucleotide probes. Login to comment 149 ABCC7 p.Gly551Asp Three of these cell clones (2b, 3c, and 6a) were highly positive for both the wild-type and the G551D genotype, as indicated by the asterisks. Login to comment 150 ABCC7 p.Gly551Asp Positive plasmid cDNA controls for the wild-type (1e) and the G551D (1f) CFTR sequences were also run as standards. Login to comment