The coding sequence of GFP in pCAG-mGFP31 was replaced with the coding sequence of HcRed from pHcRed1-1N1 (Clontech). Immunocytochemical staining was performed 48 h after transfection as described previously.32 CNG channels were detected with the anti-CNGA3 primary antibody SA3899 (1:250) ...
in silico tool unclassified variant Breast CancerPATHOGENICITYProtein Multiple Sequence Alignmentamino acid BongonsSNP AnalyzerSensitivity improvement in molecular genetic analysis has led to increased detection of novel sequence variants of unknown clinical significance in disease related genes. These ...
Sequence analysis All variants from the GPR17 gene were extracted from variant call formats of the relevant cohorts and mapped to SnpEffv4.0b annotations (51), and all nonsynonymous coding variants were mapped to dbNSFPv3.0 for functional prediction and annotation (52). Table 1 represents the fr...
These models were used to predict PSMutPred scores for each missense variant in ClinVar. Experimental materials Plasmid preparation and cell culture The full-length coding sequences of mouse Eps8 (NM_007945.4) and its mutations were PCR amplified and cloned into the pEGFP-C3 vector. The primer...
Using the coding sequence of the canonical splice isoform of MTHFR (Ensembl: ENST00000376590.9; GenBank: NM_005957.5), we produced mutagenized cDNA libraries from both reference ("wildtype," or WT) and p.Ala222Val variant templates. Each variant should be sufficiently well represented in its ...
Computational methods for estimating missense variant impact suffer from inconsistent performance across genes, which poses a major challenge for their reliable use in clinical practice. While ensemble scores leverage multiple prediction methods to enhan
Furthermore, the patient’s daughter carried this variant, whereas his wife did not carry the variant. The proband with obesity suffered eight episodes of HTG-AP from the age of 36 years, and each onset of AP was correlated to high-fat diet consumption and heavy smoking. In vitro, this ...
Genewiz Company was commissioned to synthesize the GPD1 coding sequence of wild type and p.K327N mutant, and cloned into the overexpressed plasmid vector pcDNA3.1, respectively. Sanger sequencing confirmed the accuracy of the insertion sequence, and constructed GPD1 WT and GPD1 K327N plasmids. ...
For each method, we first obtained predicted scores of all possible rare missense variants in canonical transcripts, and then sort the scores and converted the scores into rank percentile. Higher rank percentile indicates more damaging, e.g., a rank score of 0.75 indicates the missense variant is...
Through whole-genome sequencing of 15,220 Icelanders and imputation of the variants identified, we discovered a rare missense variant in NR1H4 (R436H) associating with lower levels of total cholesterol (effect = −0.47 standard deviations or −0.55 mmol L−1, p = 4.21...