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More than a week ago, the top academic journal "Science" just reported on the potential disruptive changes made by the famous scholar Professor Zhang Feng in the field of gene therapy
.
In less than half a month, Professor Zhang Feng's team published an important paper on RNA editing in the sub-Journal Nature-Biotechnology
As a pioneer of CRISPR gene editing technology, this paper by Professor Zhang Feng's team still focuses on the CRISPR-Cas13 gene editing system
.
The researchers pointed out that this Nobel Prize-winning technology has many application prospects, and one of the most cutting-edge directions is for RNA editing
If DNA editing is a blueprint for rewriting life, RNA editing is not so extreme
.
Unlike DNA editing, it does not permanently change the genetic code of the genome, but only temporarily regulates gene expression-if a disease-causing gene is overexpressed, we can control its expression through RNA editing; And if there is a mutation in a disease-causing gene, we can temporarily restore it to normal
Since RNA editing has many advantages over DNA editing, why haven't scientists made too many breakthroughs in this field before? To a large extent, this is because the CRISPR-Cas13 system is difficult to directly apply to RNA editing
.
Specifically, this is because the key Cas13 enzyme in this system is too large to be delivered by traditional AAV viral vectors-this is currently the most widely used viral vector
In order to overcome this difficulty, researchers searched for new Cas13 enzymes in the genomes of prokaryotes and viruses, and discovered two types of "super mini" Cas13 families: Cas13bt and Cas13ct
.
Compared with some Cas13 enzymes, their size is almost half the size! Because the former is more active in mammals, Cas13bt was also selected as a follow-up research object
In the small family of Cas13bt, there are a total of 16 protein members
.
Preliminary studies have shown that they have the ability to target RNA under the guidance of CRISPR RNA and "cut open" the RNA
Subsequently, the research team introduced the inactivated Cas13bt1 and Cas13bt3 proteins into the existing RNA editing tools REPAIR and RESCUE to evaluate their application potential
.
As expected, these two proteins can also perform their own functions in RNA editing tools, editing the corresponding RNA bases
▲This system can be installed in AAV2 carrier (picture source: reference [1])
What's more gratifying is that the smaller Cas13bt protein can also be directly loaded into AAV vectors for delivery
.
The researchers loaded the CRISPR RNA required by the REPAIR system together with Cas13bt1 into the AAV2 vector, confirming that it can edit human cell lines
Taken together, the Cas13bt protein discovered in this study provides an important new tool for RNA regulation in vivo
.
In cell lines, these proteins have also been conceptually verified
.
Note: The original text has been deleted
Reference materials:
[1] Kannan, S.
, Altae-Tran, H.
, Jin, X.
et al.
Compact RNA editors with small Cas13 proteins.
Nat Biotechnol (2021).
https://doi.
org/10.
1038/s41587-021- 01030-2
[2] Feng Zhang's lab develops potential breakthrough in RNA editing delivery using'ultracompact' versions of Cas13, Retrieved August 30, 2021, from https://endpts.
com/feng-zhangs-lab-develops-potential-breakthrough-in- rna-editing-delivery-with-ultracompact-versions-of-cas13/
