The latest CRISPR/Cas9 advance has made it possible to “turn on, tune in, drop out”gene. The CRISPR/Cas9 gene-editing system is not only a powerful mean of inserting new genes and knocking out genes, now it has also become a possible way to activate specific genes.
The CRISPR/Cas9 approach in the past has been demonstrated with only limited success as a way to activate specific genes, but the new CRISPR/Cas9 approach appears capable of activating genes more effectively than older approaches and may allow scientists to more easily determine the function of individual genes
The new technique was introduced last year in an online article on Nature, describing how researchers overhauled the CRISPR/Cas9 system. This study was based on their previous analysis of the structure formed when Cas9 binds to the guide RNA and its target DNA , and was improved by the use of 3D technique.
In previous efforts to revamp CRISPR/Cas9 for gene activation purposes, scientists had tried to attach the activation domains to either end of the Cas9 protein, which resulted only limited success. Later,the team realized that two small loops of the RNA guide poke out from the Cas9 complex and could be better points of attachment because they allow the activation domains to have more flexibility in recruiting transcription machinery.
Using the revamped system, the team activated about a dozen genes that had proven difficult or impossible to turn on by the previous generation of Cas9 activators. And each gene showed at least a twofold boost in transcription, and multiple orders of magnitude increase in many genes were found in activation too.
After investigating single-guide RNA targeting rules for effective transcriptional activation, the team undertaken a large-scale screen designed to identify genes with resistance to a melanoma drug called PLX-4720. They found that a gene signature based on the top screening hits correlated with a gene expression signature of BRAF inhibitor resistance in cell lines and patient-derived samples, suggesting that large-scale screens such as the one demonstrated in the current study could help researchers discover new cancer drugs that prevent tumors from becoming resistant.
This new CRISPR/Cas9 technique is of interest to researchers who are planing to use it to correct the effects of autism or neurodegenerative diseases such as Alzheimer. The CRISPR/Cas9 reagents and service have been made available to academic labs by some bio-tech suppliers such as , Creative Animodel.
Creative Animodel is a world leader in providing custom transgenic animal model service. It offers high quality CRISPR/Cas9 System for mutagenesis and genome editing, with years of experience serving hundreds of clients across six continents. Its full-service model has enabled medical and academic researchers to receive over 1,000 successful and unique gene targeted animal model lines.
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