CRISPR screens are feasible in TP53 wild-type cells

CRISPR-Cas9 technology provides an efficient method to precisely edit the mammalian genome in any cell type and represents a novel means to perform genome-wide genetic screens. A detailed protocol discussing the steps required for the successful performance of pooled genome-wide CRISPR-Cas9 screens is provided...

This manuscript describes a genome-scale cell-based screening approach to identify extracellular receptor-ligand...

This video describes wild-type blocking polymerase chain reaction, or WTB-PCR, which detects somatic mutations. The PCR-based technique uses a locked nucleic acid, or LNA, an oligonucleotide that binds to its complementary wild-type allele and blocks its elongation. The blocking selectively allows the amplification of low-level mutant alleles over their wild-type variants in the...

The article describes a protocol for the application of an in vitro model for exhaustion to complete a genome-wide CRISPR knockout screen in healthy donor chimeric antigen receptor T cells.

Source: Bennett, J. A., et al. Visual and Microscopic Evaluation of Streptomyces Developmental Mutants. J. Vis. Exp. (2018)This video demonstrates the cultivation and phenotypic comparison of wild-type and mutant Streptomyces coelicolor strains. It outlines procedures for minimizing spontaneous mutations, preventing cross-contamination, and promoting bacterial growth through proper plating techniques. Finally, the video highlights visual differences in colony morphology to illustrate the...

Source: Paschke, P., et al. Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria. J. Vis. Exp. (2019).This video demonstrates the procedure for introducing a plasmid carrying a target gene into amoeba cells via electroporation, followed by recovery in a bacterial supplement to enable efficient gene delivery in wild-type...