Fig 1. Generation of Tnnt2R141W/+ mice.
(A) Homologous recombination strategy used to introduce the DCM Arg141Trp (R141W) mutation into exon 9 of the murine Tnnt2 gene in ES cells. The resultant mice were mated with EIIa-Cre recombinase mice to remove the floxed neo/zeo cassette. (B) ES clones having undergone successful homologous recombination with the R141W targeting construct were identified by long range PCR using primer pairs F1 and R1 at the 5’ end, and F2 and R2 at the 3’ end. Positions of the primers are shown in panel A. Clones 1, 2, 4, and 5 showed appropriate homologous recombination at both the 5’ and the 3’ ends. (C) Expression of mutant Tnnt2 mRNA in Tnnt2R141W/+ mice was confirmed by reverse transcription-PCR and sequencing of total cardiac RNA. WT, wildtype.