We conducted site-directed mutagenesis (Quikchange; Roche) to generate single and double KRAS mutants (G12V, G12D, Q61H, Y32F/Q61H, and Q61H/Y64F). pCMV5-Src (WT or K295RY527F), pCMV5-SHP2 (WT and C459S), pCGN-HA-HRAS WT, pCGN-HA-KRAS WT, and pCGN-HA-NRAS WT were ...
in CaV1.3 channels has opposing effects on voltage- and Ca2+-dependent regulation Worawan B. Limpitikul, Ivy E. Dick, Manu Ben-Johny & David T. Yue† CaV1.3 channels are a major class of L-type Ca2+ channels which contribute to the rhythmicity of the heart and brain....
Cancer is a disease driven by genetic variation and mutation. Exome sequencing can be utilized for discovering these variants and mutations across hundreds of tumors. Here we present an analysis tool, VarScan 2, for the detection of somatic mutations and copy number alterations (CNAs) in exome...
To achieve accurate mutagenesis of the entire DBD (residues 102–292, 573 bp long), we generated 4 sub-libraries (labeled “A,”“B,”“C,” and “D”), covering consecutive 141–144 bp fragments of the DBD flanked by wtp53-homologues regions (Figure 1A). These sub-libraries were sepa...
Materials and methods Plant materials and growth conditions The ghd10 mutant was derived from an M2 population of the japonica rice cultivar Wuyunjing 7 (WYJ7) after EMS mutagenesis (After soaking for 24 hours, transfer the seeds into 1.5% (v/v) EMS solution at 28 degree environmental temper...
Luo D, Coen ES, Doyle S, Carpenter R (1991) Pigmentation mutants produced by transposon mutagenesis in Antirrhinum majus. Plant J 1:59–69 Article PubMed CAS Google Scholar Lyons M, Cardle L, Rostoks N, Waugh R, Flavell AJ (2008) Isolation, analysis and marker utility of novel miniatur...
Understanding the impact of guide RNA (gRNA) and genomic locus on CRISPR-Cas9 activity is crucial to design effective gene editing assays. However, it is challenging to profile Cas9 activity in the endogenous cellular environment. Here we leverage our TR
mutagenesis (Fisher’s exact test for mutated cytosines in TCG context: clone 1,P=1.1 × 10−3, OR=2.42; clone 2,P=1.0 × 10−7, OR=3.31; clone 3P=9.0 × 10−5, OR=2.28). Furthermore, C-to-G and C-to-A mutations were enriched in kataegic TCW trinucleotides (Fisher’s...
In this study, we sought to determine the role of A62V in HIV-1 viral mutagenesis and fitness in the context of clinically relevant drug-resistant mutations. To do this, we first used a single-cycle assay to assess the mutant frequency of the HIV-1 MDR complex mutants (i.e., the Q1...