Using short interfering RNA, the authors managed to decrease the RNA level of a mutant KCNH2 (hERG) allele (c.G1681A) by 61.8%, thereby increasing probability of formation of functional hERG tetramer by 4.5 times.
The genetic screening showed that the patient had the c.A2987T (p.N996I) mutation in KCNH2. In order to verify the role of this mutation in LQT syndrome development, two pairs of isogenic pluripotent cell lines that had the same genetic background and differed only by one point mutation were used.
Although iPSC-CMs express relevant ion channel genes (SCN5A, KCNJ2, CACNA1C, KCNQ1, and KCNH2), structural genes (MYH6, MYLPF, MYBPC3, DES, TNNT2, and TNNI3), and transcription factors (NKX2.5, GATA4, and GATA6) [77], they differ from adult ventricular cardiomyocytes in a number of properties.
Fukuda, "The generation of induced pluripotent stem cells from a patient with KCNH2 G603D, without LQT2 disease associated symptom," Journal of Medical and Dental Sciences, vol.
Six exons had low coverage in all samples examined by all methods [APOB (apolipoprotein B) exon 1,
KCNH2 exon 13, KCNQ1 (potassium voltage-gated channel, KQT-like subfamily, member 1) exon 1, RYR1 (ryanodine receptor 1 [skeletal]) exon 90, SCN5A (sodium channel, voltage-gated, type V, [alpha] subunit) exon 1, TGFBR1 (transforming growth factor, [beta] receptor 1) exon 1].
generated hiPSC-CMs from both symptomatic and asymptomatic patients with a G1681 mutation in
KCNH2 and put more emphasis on iPS-based disease modelling as a drug screen platform.
Here we describe a rapid single-strand conformation polymorphism-heteroduplex (SSCP-HD) analysis method for screening for mutations in all coding regions of the KCNH2 and KCNE2 genes, and identify 11 KCNH2 mutations, 6 of which were novel, and several polymorphisms in 40 families with LQTS.
Intronic PCR primer sequences were designed for robust amplification of the KCNH2 and KCNE1 exons, without nesting or additives (19, 20), under identical thermocycling conditions.
Forty probands were screened for KCNH2 and KCNE2 mutations using SSCP-HD analysis.
A rare allelic variant of KCNH2 encoding an Arg1047Leu amino acid variant was found in 3 of 80 wild-type alleles (Table 3).
We also identified eight different single-nucleotide polymorphisms (SNPs) in KCNH2 and one SNP in KCNE2 among the patients and the wild-type controls (Table 3).
The 29 probands without mutations in KCNH2 were also screened for mutations in KCNQ1, KCNE1, and SCN5A, and several additional mutations were found (M.