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As a new genetic engineering technology, gene editing is essentially to change natural selection artificially.
the widely used "gene scissors" is CRISPR-Cas9, a technology in a range of gene therapy applications have shown great potential, but large protein molecules become one of the most important bottlenecks in the future application of CRISPR.
recently, researchers at the Institute of Innovative Genomics at the University of California, Berkeley, usa, discovered a new gene editing tool, Cas, a minimally functional CRISPR-Cas system consisting of 1 to 70 KD cas protein and a CRISPR array, encoded only in the genome of giant phages, half the size of CRISPR-Cas 9, published July 16 in the journal Science.
DOI: 10.1126/science.abb1400Cas (Cas12j) is a Cas protein family encoded in the branches of giant phages, which are used by giant phages to induce bacteria to resist themselves rather than their own viruses.
it uses a single active site for CRISPR RNA (crRNA) processing and crRNA-directed DNA cutting to target foreign nucleic acids.
researchers first determine whether Cas can be used as a true CRISPR-Cas system.
they found that Cass is a functional phage protein and a true CRISPR-Cas effector in its natural environment, which can elicit the complementary DNA of crRNA.
, this single RNA system is much more compact than other active CRISPR-Cas systems.
next, the team needed to understand cassin's in vitro DNA recognition and cleavage requirements.
results show that the lysis activity of Cas can only be observed when the trace DNA is identified, and only the minimum PAM condition is required, which may facilitate broader nucleic acid testing.
finally, to study whether Cas can be used in human genome editing, the researchers used Cass, which is expressed with crRNA, in heK293 cells to perform gene destruction determinations.
they found that Cas-2 and Cas-3 induced the targeted destruction of genomic-integrated-enhanced green fluorescence protein (EGFP) genes.
, Cas-2 with a single guided RNA is able to edit up to 33% of the cells, which is comparable to the previously reported levels of CRISPR-Cas9, CRISPR-Cas12a and CRISPR-CasX.
the macrophage-coded Cas-in-the-host's functional diagram in the case of its host's hyper-infection actually, this is not the first appearance of Cas.
it was first discovered by Basem Al-Shayeb, a doctoral student at IGI, another lead author of the study, published in Nature on February 12.
, Al-Shayeb was working in a lab at the University of California, Berkeley, where Heltian F. Banfield, a professor in the Department of Earth and Planetary Sciences, worked.
they believe that the giant phage, which contains Cas, is a member of the giant phage that already exists and exists in a variety of environments.
recent discovery reveals cassic potential for gene editing, providing an advantage for cell transfer and expanding the "toolbox" of gene editing.
more important, Cass protein brings giant phages to the forefront of human health discovery and biotechnology applications.
but researchers have a long way to go to optimize Cas for gene editing and to explore the best way to design guidance RNAs for targeting specific genes.
References: crispR-CasF from huge phages is a hypercompact genome case.