Introduction to CRISPR Functional Research Guide

Genome sequencing has made us aware that only a small portion of the human genome is translated into proteins. In fact, the 80% of our genome will be transcribed into RNA, but most of these transcripts do not produce protein. In recent years, people have found that non coding RNA is often associated with human diseases, but the vast majority of non coding RNA function is still unknown. CRISPR Cas9 can play an important role in this area.

CRISPR activation (CRISPRa) and CRISPR inhibition (CRISPRi) are particularly suitable for the analysis of the specific features of non coding RNA. Although more and more researchers begin using CRISPRa and CRISPRi, they are just starting.

CRISPRa

Catalyze inactivation of Cas9 (dCas9) and link the transcriptional activator, which can effectively promote the transcription of the genome specific location.

cDNA overexpression is a commonly used method to increase the expression level of individual genes. Compared with this method, CRISPRa is simpler to use, and can activate multiple genes simultaneously. Previous researchers have linked the domain of activated transcription to the dCas9's terminus, but this system is often unable to initiate gene transcription. Two loop RNA protrudes from the cas9 complex is the best linkage sites. such activation domain is more flexible in the recruitment of the transcriptional machinery.

The closer to the transcription start site, the stronger the effect of transcriptional activation. If the CRISPRa is targeted to the upstream (more than 200bp) of the transcription initiation site, the transcription will be more temperate and closer to the physiological level.

Research team integrated a series of short peptides (that is, SunTag array) in the dCas9. These short peptides are equivalent to a set of molecules linked to the recruitment of multiple copies of the transcription activator in the cell, to generate strong signals.

Which version of CRISPRa will eventually come to the fore? Experts believe it is too early to come to the conclusion. There is no harm in trying out several ways in their own research area, and the better way may also be about to come out.

Company :-CD Genomics

User :- Debbie Evans

Email :-wangyizhusss@gmail.com

Phone :---

Mobile:- -

Url :- http://www.cd-genomics.com/Custom-Libraries.html