Htr2c^tm1.1Eme

Overlap-extension PCR replaced 5 adenosines in the editing sites within exon 5 with guanosines, which mimics the base-pairing properties of inosine. A floxed neo cassette was inserted downstream of exon 5 and removed by cre mediated recombination.The predicted product represents a fully edited (VGV) isoform. RNase protection assay confirmed the decreased expression of a splice variant that uses the more proximal donor site in exon 5 (RNA1) and the increased expression of a splice isoform encoding the full length receptor protein. Expression of the splice variant that uses the distal 5' splice site in intron 5 (RNA3) is unchanged. Western blot analysis confirmed increased protein expression in the brain. (J:163043)