Category Ranking
98%
Total Visits
921
Avg Visit Duration
2 minutes
Citations
20
Article Abstract
In this protocol, we take CRISPR/Cas9 and Gal4/UAS approaches to achieve tissue-specific knockout in parallel with rescue of the knockout by cDNA expression in . We demonstrate that guide RNAs targeting the exon-intron junction of target genes cleave the genomic locus of the genes, but not transgenes, in a tissue where Gal4 drives Cas9 expression. The efficiency of this approach enables the determination of pathogenicity of disease-associated variants in human genes in a tissue-specific manner in . For complete details on the use and execution of this protocol, please refer to Yap et al. (2021).
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9204798 | PMC |
| http://dx.doi.org/10.1016/j.xpro.2022.101465 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Transcriptional Readthrough at Locus Suppresses and Impairs Heart Development.
bioRxiv
August 2025
Department of Medicine, University of California San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA.
Background: Activating Transcription Factor 4 (ATF4) functions as a transcriptional regulator in various cell types and tissues under both physiological and pathological conditions. While previous studies have linked ATF4 activation with promoting cardiomyocyte (CM) death in dilated cardiomyopathy (DCM), atrial fibrillation, and heart failure, its role in developing CMs remains unexplored.
Methods: We generated multiple distinct CM-specific ( , and ) and global knockout ( and ) mouse models targeting different regions, as well as cardiomyocyte-specific deletion of to study cardiac phenotypes.
Adipocyte-specific IGF1R knockout activates the β-catenin/apelin axis to combat diet-induced obesity in male mice.
Diabetes Obes Metab
September 2025
Institute of Genome Engineered Animal Models for Human Diseases, National Center of Genetically Engineered Animal Models for International Research, Dalian Medical University, Dalian, China.
Aims: Obesity, driven by complex genetic and environmental interactions, remains a global health crisis with limited therapeutic options. The insulin-like growth factor 1 receptor (IGF1R) plays dual roles in metabolism and growth, but its tissue-specific functions in adipose biology are controversial. This study investigates how adipose-specific IGF1R knockout impacts systemic metabolism under high-fat diet (HFD) stress and explores the underlying mechanisms.
View Article and Find Full Text PDFT cell immunity depends on the precise coordination of signaling networks with actin cytoskeleton remodeling, yet the molecular regulators of these processes remain incompletely defined. Flightless-1 (FLII) is a gelsolin-family actin regulator with unique leucine-rich repeats that can couple cytoskeletal dynamics to diverse signaling pathways. Here, using conditional knockout mice, we identify essential roles for FLII in both CD8⁺ and regulatory T cells.
View Article and Find Full Text PDFKnockout Mice as an Experimental Model to Study the Biology of Inositol Pyrophosphates.
Methods Mol Biol
August 2025
Laboratory of Cell Signalling, Centre for DNA Fingerprinting and Diagnostics, Hyderabad, India.
Knockout mice have served as excellent model systems to investigate the functions of specific mammalian proteins at the organismal level. Several studies on tissue-specific or whole-body depletion of individual IP6 kinase (IP6K) paralogs have shed light on myriad roles for their product, the inositol pyrophosphate 5-InsP, in different physiological processes and pathological states. The loss of Ip6k1, Ip6k2, or Ip6k3 leads predominantly to nonoverlapping phenotypes in mice, reflecting their differential tissue, cell type, and subcellular distribution.
View Article and Find Full Text PDFGenetic ablation of Pth4 disrupts calcium-phosphate balance, bone development, and kidney transcriptome in teleosts.
Gen Comp Endocrinol
August 2025
Acuabiotec Lab. Institute of Marine Research, Spanish National Research Council (IIM-CSIC), 36208 Vigo, Spain. Electronic address:
Parathyroid hormone 4 (Pth4) is an evolutionarily conserved member of the PTH family, expressed in hypothalamic neurons and lost in eutherian mammals. In order to elucidate its role in mineral homeostasis and skeletal development, a pth4 knockout (pth4) zebrafish line was generated using CRISPR/Cas9 and transcriptomic profiling was conducted across six key tissues: brain, kidney, intestine, gills, scales, and bone. The results obtained demonstrated that the loss of Pth4 led to pronounced disturbances in calcium and phosphate homeostasis, skeletal deformities, and widespread tissue-specific transcriptional alterations.
View Article and Find Full Text PDF