In the rice blast fungusMagnaporthe oryzae, a CRISPR-Cas9 genome editing system based on expression of Cas9 and CRISPR componentsin vivohas been reported to target single genes14. However, the generation of mutants using this procedure has not been widely adopted and the protocol requires labour-i...
The functional mechanism of CRISPR/Cas9 has been gradually revealed, and natural CRISPR/Cas9 has been rapidly applied to bacterial transformation.44,53In 2011, Siksnys et al. transferred the first CRISPR gene sequence fromStreptococcus thermophilustoE. coli, and theE. colithat received the CRISPR ge...
2 基于CRISPR/Cas9技术的遗传性血液病基因治疗策略 CRISPR/Cas9系统由向导RNA (single-guide RNA, sgRNA)和Cas9蛋白组成[17,18,19],与ZFN和TALEN技术类似,CRISPR/Cas9技术同样通过诱导DNA双链断裂,诱发同源重组修复(homology- directed repair, HDR)和/或非同源末端连接(non- homologous end joining, NHEJ)等DNA损...
CRISPR/Cas9 mediated gene knockout is a powerful tool for genome editing with the ability to target multiple genes simultaneously. Establishing an efficient, multiplexed gene knockout system using CRISPR/Cas9 that is both simple and robust in its applica
CRISPR-Cas9 has been developed as a therapeutic agent for various infectious and genetic diseases. In many clinically relevant applications, constitutively active CRISPR-Cas9 is delivered into human cells without a temporal control system. Excessive and
Brain tumours are the leading cause of cancer-related death in children, and there is no effective treatment. A growing body of evidence points to deregulated epigenetics as a tumour driver, particularly in paediatric cancers as they have relatively few
Multiplexed CRISPR-Cas9 gene-editing is efficient in primary human T cells. a Schematic representation of the EGFRvIII targeted CAR construct. b Primary human T cells were stimulated, RNP electroporated and transduced to produce CAR T cells. c Following expansion, cells were subjected to flow cyto...
536–2–13–16 retained the 2 bp mutation at the CRISPR target site while plant WPT536–2–13–15 segregated back to homozygosity for the wild-type allele. Given these findings, it was determined that plant WPT536–2–13–16 is a simple M2 generation plant (contains a mutation, but ...
(WT) larvae (left), indicative of the presence of thenatterin-liketranscripts; and in the depleted (KO) larvae by CRISPR/Cas9 system (crRNA natterin-like WD07479944, Sigma) (right). The embryos body changed the shape due to long-lasting high temperature incubation in conical-bottom ...
We then adopted the CRISPR-Cas9 system to specifically silence CD44 at the DNA level in human highly metastatic osteosarcoma cells (MNNG/HOS and 143B), and further determined the effects of CD44 knockout on function in human highly metastatic osteosarcoma cells. Materials and Methods Meta-analysis...