Microbial CRISPR-Cas defense systems have been adapted as a platform for genome editing applications built around the RNA-guided effector nucleases, such as Cas9. We recently reported the characterization of Cpf1, the effector nuclease of a novel type V-A CRISPR system, and demonstrated that it ...
Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins RNA-guided engineered nucleases (RGENs) derived from the prokaryotic adaptive immune system known as CRISPR (clustered, regularly interspaced, short palind... S Kim,D Kim,SW Cho,... - 《Gen...
Among all the CRISPR/Cas9 systems, Type II CRISPR/Cas9 has a comparatively more straightforward structure, broadening its genetic engineering applications. It consists of three vital components: crRNA, tracrRNA combined to form a sgRNA andStreptococcus pyogenesoriginated-Cas9 protein. The sgRNA is create...
It has been reported that the efficiency of genome editing through HDR is influenced by chemical or genetic disruption of the NHEJ pathway15,16. The efficiency of HDR can also be increased by controlling the timing of the delivery of SpyCas9-single guide RNA (sgRNA) ribonucleoprotein (RNP) ...
通过在倒位片段的两端设计向导RNA,引导Cas9切割倒位片段两端,诱发非等位同源重组,使倒位片段再次发生倒位,实现患者iPSC中内含子倒位的修复,基因修复的细胞分化后凝血因子VIII基因表达量由0上升至正常水平的50%~120%,但修复效率只有3.7%[36]。这可能由于细胞中存在一系列抑制非等位同源重组的调控机制[40](表3)。
In most cases, researchers rely on viruses to carry the gene for Cas9, as well as the RNA guide strand. In 2014, Anderson, Yin, and their colleagues developed a nonviral delivery system in the first-ever demonstration of curing a disease (the liver disorder tyrosinemia) with CRISPR in an...
Beyond genome editing, CRISPR/Cas can be used as a platform for RNA guided DNA–protein interactions, and thereby deliver various effector domains to a specific genomic location. By introducing point mutations into the two nucleolytic domains, nuclease deficient versions of Cas9, called dead Cas9 ...
CRISPR/Cas9, CRISPR/Cas12 and CRISPR/Cas13. CRISPR-associated protein 9 (Cas9), derived from the CRISPR Type II bacterial immune system, was an RNA-guided DNA endonuclease that could program for new sites by changing the sequence of its guided RNA [48]. It was developed as a tool to ...
Simple and efficient gene editing using the RNA-guided nuclease CRISPR/Cas9 The last two years have seen the development of a new approach to build endonucleases with customized sequence specificities, which has revolutionized gene editing by its simplicity and efficiency. The approach is borrowed from...
M. RNA-Guided Human Genome Engineering via Cas9. Science 2013, 339 (6121), 823. Misteli, T.; Spector, D. L. Applications of the Green Fluorescent Protein in Cell Biology and Biotechnology. Nat Biotech 1997, 15 (10), 961-964. Morgan, S. L.; Mariano, N. C.; Bermudez, A.; ...