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Slow channel congenital myasthenic syndrome (SCCMS) is a disorder of the neuromuscular synapse caused by dominantly inherited missense mutations in genes that encode the muscle acetylcholine receptor (AChR) subunits. Here we investigate the potential of post-transcriptional gene silencing using RNA interference (RNAi) for the selective down-regulation of pathogenic mutant AChR. By transfection of both siRNA and shRNA into mammalian cells expressing wild-type or mutant AChR subunits, we show, using 125I-alpha-bungarotoxin binding and immunofluorescence to measure cell surface AChR expression, efficient discrimination between the silencing of alphaS226F AChR mutant RNA transcripts and the wild-type. In this model we find that selectivity between mutant and wild-type transcripts is optimized with the nucleotide mismatch at position 9 in the shRNA complementary sequence. We also find that allele-specific silencing using shRNA has comparable efficiency to that using siRNA, underlining the general potential of stable expression of shRNA molecules as a long term therapeutic approach for allele-specific silencing of mutant transcripts in dominant genetic disorders.

Original publication




Journal article


Hum Mol Genet

Publication Date





2637 - 2644


Alleles, Animals, Base Pair Mismatch, Base Sequence, Cell Line, DNA, Down-Regulation, Gene Silencing, Humans, Microscopy, Fluorescence, Molecular Sequence Data, Mutation, Oligonucleotides, Promoter Regions, Genetic, RNA, RNA Interference, RNA, Small Interfering, Receptors, Cholinergic, Synapses, Transcription, Genetic, Transfection