2023年7月20日 · Homology-directed repair (HDR), a method for repair of DNA double-stranded breaks can be leveraged for the precise introduction of mutations supplied by synthetic...

MMEJ作为替代末端连接的一种形式，只需要非常小的同源区域 (5~25bp)进行修复，使得构建靶向载体更加容易。 Takashi Yamamoto实验室利用MMEJ创建了CRISPR基因敲入的新方法，命名为PITCh (精确整合到目标染色体中)。 利用它们的PITCh质粒，可以在大约一个半月的时间内创建GFP敲入细胞系，而不需要进行复杂的同源臂克隆。 DNA双链断裂后的修复方法主要有3种。 HDR从具有两侧序列同源性的修复模板中复制该序列，用于精确的DSB修复。 NHEJ以易错的 …

2019年12月2日 · Broadly speaking, DNA DSBs are reversed in two ways: re-ligation of the ends through pathways such as non-homologous end joining (NHEJ) and microhomology-mediated end joining (MMEJ) that can...

2024年8月10日 · By combining with an inhibitor of non-homologous end joining (NHEJ) or the HDRobust strategy, these modular ssDNA donors achieve up to 90.03% (median 74.81%) HDR efficiency. The HDR-boosting...

2023年6月23日 · The efficiency of homology-directed repair (HDR) plays a crucial role in the development of animal models and gene therapy. We demonstrate that microhomology-mediated end-joining (MMEJ) constitutes a substantial proportion of …

2024年4月29日 · We demonstrate that the MMEJ pathway plays an important role in CRISPR-Cas9-induced LD and find two key MMEJ genes (POLQ and RPA) perform oppositely in CRISPR-Cas9-induced LD and HDR. We provide a potentially safe strategy to decrease the CRISPR-Cas9-induced LD and increase HDR efficiency by modulating POLQ and RPA.

2021年7月1日 · End resection directs DSB repair through template-independent microhomology-mediated end joining (MMEJ) or template-dependent homology-directed repair (HDR). For MMEJ, small homologous DNA sequences (5–25 bp) within the two resected ends are paired and lead to DNA repair resulting in small deletions or insertions.

2018年6月1日 · Common genome-editing strategies are either based on non-homologous end joining (NHEJ) or, in the presence of a template DNA, based on homologous recombination with long (homology-directed repair [HDR]) or short (microhomology-mediated end joining [MMEJ]) homologous sequences.

2018年7月7日 · We compared the efficiencies between HDR- and MMEJ-mediated targeted substitutions in the mVenus-off reporter cell line. For HDR, we prepared one efficient gRNA that cuts near the 2X stop codon site of the mVenus -off sequence and a 200-base ssODN as the HDR donor template.

2022年2月25日 · Compared to the single-stranded oligo deoxyribonucleotide (ssODN)-mediated homology directed repair (HDR), LAHR yields relatively high editing efficiency as demonstrated for both a reporter gene and endogenous genes. We found that both HDR and microhomology-mediated end joining (MMEJ) mechanisms are involved in the LAHR process.