A simple method using CRISPR-Cas9 to knock-out genes in murine ... - Nature
WEBDec 18, 2020 · Gene knock-out (KO) is a critical method for identifying the functions of coding and non-coding genomic regions. Recent emergence of genome editing tools such as CRISPR/Cas9 can be used to...
CRISPR Guide - Addgene
WEBLearn how CRISPR systems work and how to use them for genome engineering. Find out how to design and generate gRNAs, nickases, and high fidelity Cas9s for various applications.
Guidelines for optimized gene knockout using CRISPR/Cas9
WEBHere, we present guidelines and tools to optimize CRISPR/Cas9 genome-targeting efficiency and specificity. The nature of the target locus, the design of the single guide RNA and the choice of the delivery method should all be carefully considered prior to a genome-editing experiment.
An efficient and precise method for generating knockout cell lines ...
WEBSep 7, 2020 · Here, we adapted a targeted knock‐in strategy with CRISPR‐Cas9 system and characterized the efficiency for generating single or double knockout cell lines. Specifically, a homology‐arm based donor cassette consisting of genes encoding a fluorescence protein and antibiotic selection marker driven by a …
An improved strategy for CRISPR/Cas9 gene knockout and …
WEBFeb 13, 2020 · The gene knockout (KO) efficiency of CRISPR/Cas9 mainly depends on cellular Non-homologous End Joint (NHEJ) repair to maintain genomic integrity if a complementary donor is not present when Cas9 cleaves both double strands at …
Genome-scale CRISPR-Cas9 knockout and transcriptional
WEBDec 10, 2014 · Here we describe a protocol for genome-scale knockout and transcriptional activation screening using the CRISPR-Cas9 system. Custom- or ready-made guide RNA libraries are constructed and...
Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing
WEBAug 21, 2021 · Clustered regularly interspaced short palindromic repeat (CRISPR) and their associated protein (Cas-9) is the most effective, efficient, and accurate method of genome editing tool in all living cells and utilized in many applied disciplines.
CRISPR in cancer biology and therapy - Nature Reviews Cancer
WEBFeb 22, 2022 · This paper describes the development of a ‘PAMless’ CRISPR–Cas9 variant, SpRY, that dramatically broadens the number of possible genomic targets for nuclease- or non-nuclease-mediated editing.
Protocol for generating monoclonal CRISPR-Cas9-mediated knockout …
WEBSep 15, 2023 · CRISPR-Cas9 is a powerful technology for accurate and optimizable genome editing. Here, we present a protocol for generating monoclonal knockout (KO) cell lines using CRISPR-Cas9, ribonucleoprotein complexes (RNPs), and lipofection in adherent HNSCC cells from start to finish.
Simplified Gene Knockout by CRISPR-Cas9-Induced Homologous ...
WEBDec 9, 2021 · Here we designed and characterized a knockout fragment intended to repair Cas9-induced gene disruptions by homologous recombination. We identified knockout clones of Komagataella phaffii with high fidelity by …