Breakthrough Technology to Improve the Range and Accuracy of Cas9 Targ

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Breakthrough Technology to Improve the Range and Accuracy of Cas9 Target:

Breakthrough Technology to Improve the Range and Accuracy of Cas9 Target

Slide2:

Scot A Wolfe, an associate professor of molecular, cell and cancer biology at the University of Massachusetts School of medicine, reported that they developed a new technology that could significantly improve the accuracy of Cas9 and expand its target range by fusing the DNA binding domain (pDBD) and CRISPR/Cas9 System . The research was published on October 19th in the Nature Methods.

Slide3:

Because of its easy use and efficiency, the CRISPR-Cas9 genome editing system is completely changing the biological sciences. The most commonly studied Cas9 nucleic acid enzyme (SpCas9) is derived from Streptococcus pneumoniae. SpCas9 and its related RNA only will be issued to a section of the DNA sequence based on the two stages of sequence review.

Slide4:

Since it is a very important factor to restrict the Cas9 target, the Cas9 element is a key factor to the PAM, which is a complementary single oriented RNA (sgRNA). The simplicity of the SpCas9-sgRNA system makes it easy to edit the genomes of various organisms and cell lines.

Slide5:

The accuracy of SpCas9 has not yet reached the best standards for most genes involved in editing a large number of cells. Many studies have confirmed that SpCas9 can be in the non - expected site of cutting genes. SpCas9 accuracy analysis of the whole genome engineering using the crispr cas9 system in the near future shows that there are only two nucleotide differences between the majority of the genomic sites and the RNA sequence, and a part of the genome has three nucleotides with a mismatch.

Slide6:

In this Nature Methods article, researchers at the University of Massachusetts Medical School confirmed and reduced the cas9 intrinsic DNA binding affinity by integrating such as Cys2-His2 zinc finger proteins (ZFPs) or transcription activation sample effect factor (TALEs). They were able to generate a Cas9-pDBD chimera, greatly improving cas9 accuracy and increasing the targeting range. Since it could easily control the specificity and affinity of this platform, Cas9-pDBDs provided us with a flexible and customizable system to achieve extremely accurate genome editing in which almost all of the genomic loci.

Slide7:

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