It is important to know that cells possess two major repair pathways – Non-Homologous End Joining (NHEJ) and Homology Directed Repair (HDR) – and how these pathways work, as this could be relevant when planning your experiment. Here, we’ll discuss NHEJ, and how it impacts what happens to Cas9-mediated DSBs in the genome.

May 19, 2017 · Here we devise a homology-mediated end joining (HMEJ)-based strategy, using CRISPR/Cas9-mediated cleavage of both transgene donor vector that contains guide RNA target sites and ∼ 800 bp of...

Nov 24, 2016 · Here, we describe a CRISPR-Cas9 assisted non-homologous end-joining (CA-NHEJ) strategy for the rapid and efficient inactivation of bacterial gene (s) in a homologous...

Non-homologous end joining (NHEJ), and homology-directed repair (HDR) pathways are the two mechanisms to repair DSBs created by Cas-9 protein in CRISPR/Cas-9 mechanism. 16 NHEJ facilitates the repair of DSBs by joining DNA fragments through an enzymatic process in the absence of exogenous homologous DNA and is active in all phases of the cell ...

Oct 19, 2018 · NHEJ is inherently accurate in repair of Cas9-induced DNA double strand breaks and can be harnessed to improve CRISPR/Cas9 genome editing requiring precise deletion of a defined length.

Oct 19, 2018 · Knowledge of accurate NHEJ in repair of Cas9-induced DSBs could help yield a strategy to shift NHEJ away from accurate NHEJ or even to harness accurate NHEJ to generate precise indels of defined length, promoting CRISPR/Cas9-mediated gene knockouts.

Jun 15, 2021 · Genome editing using CRISPR-Cas9 nucleases is based on the repair of the DNA double-strand break (DSB). In eukaryotic cells, DSBs are rejoined through homology-directed repair (HDR), non-homologous end joining (NHEJ) or microhomology-mediated end joining (MMEJ) pathways.

Nov 7, 2019 · Non-homologous end-joining (NHEJ) is the preferred mechanism used by hematopoietic stem cells (HSCs) to repair double-stranded DNA breaks and is particularly increased in cells deficient in the Fanconi anemia (FA) pathway. Here, we show feasible correction of compromised functional phenotypes in hem …

In this study, we report a new powerful method of generating plasmid-based KI in mice and rats using the CRISPR-Cas9 system. The principle concept of this method is the combination of highly efficient editing via NHEJ and the low efficiency, but precise editing of HDR in zygotes.

Dec 19, 2016 · Suzuki et al. developed homology-independent targeted integration (HITI) for CRISPR–Cas9- and NHEJ-based gene knock-in. HITI successfully mediated site-directed GFP knock-in in postmitotic...