Category Ranking
98%
Total Visits
921
Avg Visit Duration
2 minutes
Citations
20
Article Abstract
Allelic series are extremely valuable genetic tools to study gene function and identify essential structural features of gene products. In mice, allelic series have been engineered using conventional gene targeting in embryonic stem cells or chemical mutagenesis. While these approaches have provided valuable information about the function of genes, they remain cumbersome. Modern approaches such as CRISPR-Cas9 technologies now allow for the precise and cost-effective generation of mouse models with specific mutations, facilitating the development of allelic series. Here, we describe procedures for the generation of three types of mutations used to dissect protein function in vivo using CRISPR-Cas9 technology. This step-by-step protocol describes the generation of missense mutations, large in-frame deletions, and insertions of genetic material using SCY1-like 1 (Scyl1) as a model gene.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/bs.mie.2022.03.053 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Mutant huntingtin exon 1 protein detected in mouse brain with neoepitope antibody: effects of CAG repeat expansion, MutS Homolog 3 silencing and aggregation.
Brain Commun
August 2025
Department of Neurology, Massachusetts General Hospital, Charlestown, MA 02129, USA.
was identified in human and mouse Huntington's disease brain as the pathogenic exon 1 mRNA generated from aberrant splicing between exon 1 and 2 of that contributes to aggregate formation and neuronal dysfunction. Detection of the huntingtin exon 1 protein (HTT1a) has been accomplished with Meso Scale Discovery, Homogeneous Time Resolved Fluorescence and immunoprecipitation assays in Huntington's disease knock-in mice, but direct detection in homogenates by gel electrophoresis and western blot assay has been lacking. Subcellular fractions prepared from mouse and human Huntington's disease brain were separated by gel electrophoresis and probed by western blot with neoepitope monoclonal antibodies 1B12 and 11G2 directed to the C-terminal eight residues of HTT1a.
View Article and Find Full Text PDFCombining evidence from human genetic and functional screens to identify pathways altering obesity and fat distribution.
Am J Hum Genet
August 2025
Big Data Institute, Li Ka Shing Centre for Health Information and Discovery, University of Oxford, Oxford, UK; Centre for Human Genetics, Nuffield Department of Medicine, University of Oxford, Oxford, UK; Nuffield Department of Women's and Reproductive Health, Medical Sciences Division, University o
Overall adiposity and body fat distribution are heritable traits associated with altered risk of cardiometabolic disease and mortality. Performing rare-variant (minor allele frequency <1%) association testing using exome-sequencing data from 402,375 participants of European ancestry in the UK Biobank for nine overall and tissue-specific fat distribution traits, we identified 19 genes where putatively damaging rare variation associated with at least one trait (Bonferroni-adjusted p < 1.58 × 10) and 50 additional genes at false discovery rate (FDR) ≤1% (p ≤ 4.
View Article and Find Full Text PDFAssessment of hepatitis C virus permissiveness in iteratively genetically humanized mice.
J Virol
September 2025
Department of Molecular Biology, Princeton University, Princeton, New Jersey, USA.
Hepatitis C virus (HCV) is an enveloped, positive-sense single-stranded RNA virus causing chronic infections in over 50 million people who are at risk of developing severe liver disease. Greater understanding of HCV pathogenesis and vaccine development has been hampered by the lack of a fully immunocompetent small-animal model permissive to infection. Rodents are resistant to HCV infection due to a variety of factors at the levels of entry and replication, many of which have been discovered within the past decade.
View Article and Find Full Text PDFDouble non-allelic somatic activating oncogene variants in a series of mosaic vascular anomalies.
Br J Dermatol
September 2025
Université Bourgogne Europe, INSERM, CTM UMR1231, Equipe GAD, Dijon, France.
Bi-Allelic Variants in MICU1 Cause Myopathy With Extrapyramidal Signs: Case Series, Phenotypic Spectrum, and Genotype-Phenotype Correlations From 61 Patients.
Clin Genet
September 2025
Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Myopathy with extrapyramidal signs (MPXPS) is a rare, autosomal-recessive, multisystem disorder caused by biallelic loss-of-function (LOF) variants in MICU1, the calcium-sensing gatekeeper of the mitochondrial calcium uniporter. We clinically and genetically characterized seven affected individuals from six Iranian-Turkish consanguineous families and combined these data with 54 previously published cases (total of 62). The targeted neuromuscular assessment, along with muscle biopsy and exome sequencing, identified six pathogenic MICU1 variants, including c.
View Article and Find Full Text PDF