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| http://dx.doi.org/10.1111/pbi.13601 | DOI Listing |
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Molecular Characterization, Expression Analysis, and CRISPR/Cas9 Mediated Gene Disruption of Myogenic Regulatory Factor 4 (MRF4) in Nile Tilapia.
Curr Issues Mol Biol
December 2024
Department of Fisheries Science, Chonnam National University, Yeosu 59626, Republic of Korea.
Myogenic regulator factors (MRFs) are essential for skeletal muscle development in vertebrates, including fish. This study aimed to characterize the role of () in muscle development in Nile tilapia by cloning from muscle tissues. To explore the function of , CRISPR/Cas9 gene editing was employed.
View Article and Find Full Text PDFCRISPR-Cas9 Mediated Gene Deletion in Human Pluripotent Stem Cells Cultured Under Feeder-Free Conditions.
J Vis Exp
November 2024
Department of Biochemistry and Molecular Biology, College of Medicine and Health Sciences, United Arab Emirates University; Zayed Center for Health Sciences, United Arab Emirates University; ASPIRE Precision Medicine Research Institute Abu Dhabi (PMRI-AD), United Arab Emirates University;
The CRISPR-Cas9 system for genome editing has revolutionized gene function studies in mammalian cells, including stem cells. However, the practical application of this technique, particularly in pluripotent stem cells, presents certain challenges, such as being time- and labor-intensive and having low editing efficiency. Here, we describe the generation of a CRISPR-mediated gene knockout in a human embryonic stem cell (hESC) line stably expressing sgRNAs for the L2HGDH gene, using a highly efficient and stable lentiviral-mediated gene delivery system.
View Article and Find Full Text PDFEfficient CRISPR/Cas9 Knock-in Approaches for Manipulation of Endogenous Genes in Human B Lymphoma Cells.
Curr Protoc
November 2024
Division of Infectious Diseases, Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts.
Article Synopsis
- Traditional overexpression techniques struggle to accurately analyze protein-protein interactions and expression in B lymphocytes due to their resistance to lipid transfection.
- The article discusses advanced CRISPR/Cas9 knock-in methods that increase efficiency in tagging endogenous proteins, along with protocols for optimizing cutting efficiency and sgRNA selection.
- Detailed methodologies for engineering B lymphoma cells are provided, including assessing editing efficiency, designing repair templates, electroporation, and selecting engineered cells, enabling broader application in other cell types.
Expression of the C-allele of intronic rs8192675 in SLC2A2 is associated with improved glucose response to metformin.
Genet Mol Biol
September 2024
Affiliated Hospital 2 of Nantong University, Department of Radiology, No.666 Shengli Road, Nantong, Jiangsu Province, China.
Glucose is a critical nutrient for energy metabolism. The SLC2A2 gene is essential for glucose sensing and homeostasis, as it encodes the facilitated glucose transporter GLUT2. During diabetes treatment, the C-allele of rs8192675 in SLC2A2 has been found to regulate the action of metformin and reduce the absolute level of HbA1c more effectively than the T-allele.
View Article and Find Full Text PDFFast Fission Yeast Genome Editing by CRISPR/Cas9 Using Gap Repair and Fluoride Selection.
Methods Mol Biol
November 2024
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.
We present a protocol to perform CRISPR/Cas9-mediated genome editing in the fission yeast Schizosaccharomyces pombe that does not require cloning and uses the fluoride exporter channel Fex1 as the selection marker. Transformation is typically carried out on the same day of PCR primer arrival and successfully edited strains are selected 5 days after transformation. We expect the adoption of this protocol to further accelerate the throughput of genome editing in S.
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