2023年3月30日 · Two SpCas9 variants, namely, nCas9 (D10A) and nCas9 (H840A), which cleave target (guide RNA-pairing) and non-target DNA strands, respectively, are widely used for various purposes, including...

2024年6月17日 · Base editors (BEs) are a recent generation of genome editing tools that couple a cytidine or adenosine deaminase activity to a catalytically impaired Cas9 moiety (nCas9) to enable specific base conversions at the targeted genomic loci.

CRISPRCas9技术的潜在脱靶效应可能是由Cas9核酸酶活性引起的。 为了改善由Cas9核酸酶活性引发的脱靶效应，开发了Cas9 Nickase。 Cas9 Nickase是Cas9的突变形式， 是由RuvC1和 HNH两个核酸酶活性区域中其中一个核酸酶活性区域突变而成。 这种 突变形式产生单链缺口而不是在靶位点的双链断裂。 利用Cas9 Nickase进行基因组编辑，需要两个gRNA而不是一个。 两个gRNA将被设计在相反的DNA链上且紧密接近（序列相距不超过20bp），以确保一旦两条链 …

2025年4月9日 · Takesue et al. introduce BITREx, a novel genome editing method that strategically directs a single Cas9 nickase adjacent to a target tandem gene array, enabling its efficient expansion. By leveraging replication fork breakage, BITREx drives copy-number amplification, offering a unique approach to genome manipulation.

2024年8月14日 · Based on these considerations, we have conceived a strategy to induce SDs via nCas9-mediated paired nicking. Our strategy involves positioning two nCas9s upstream and downstream of a replication origin. One nCas9 targets the …

Nishimasu et al. developed an engineered Cas9, Streptococcus pyogenes Cas9 (SpCas9)-NG, a variant of Cas9 that can recognize NG-PAM instead of NGG-PAM, to expand the targeting range and improve its compatibility to the target genomic loci.

2 天之前 · Autophagy boosts HR-associated gene editing efficiency with nCas9 or enzymatically inactive Cas9 but does not affect the single-stranded template repair (SSTR) strategy. (A) Targeted deep sequencing analysis on the indicated target genes. HEK293T cells were transfected with an empty vector or Cas9 mutants with an sgRNA and 3xHA (hemagglutinin ...

Keywords: CRISPR‐Cas9, gene editing, ... (EcKlenow) fused with nCas9. In addition, they also contain an ssDNA recruitment domain, HUH endonucleases (HUHes) tethered to nickase. The second component in this system is a click DNA (clk DNA) which is a single‐stranded DNA, containing recognition sequences for binding with HUHe, ...

2016年4月15日 · Here, we describe a Cas9 D10A -based screening approach that combines an All-in-One Cas9 D10A nickase vector with fluorescence-activated cell sorting enrichment followed by high-throughput...

2020年7月15日 · Fusing a deaminase with the Cas9 nickase (nCas9) forms cytosine base editors (CBEs), which enable programmable conversion of cytidine-to-thymidine (C-to-T) mutations within a specific region of the genomic DNA without causing double-stranded breaks (1–3).