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CRISPR interference (CRISPRi) is a genetic perturbation technique that allows for sequence-specific repression of gene expression in prokaryotic and eukaryotic cells. [1] It was first developed by Stanley Qi and colleagues in the laboratories of Wendell Lim , Adam Arkin, Jonathan Weissman , and Jennifer Doudna . [ 2 ]
CRISPR-Cas9 genome editing techniques have many potential applications. The use of the CRISPR-Cas9-gRNA complex for genome editing [10] was the AAAS's choice for Breakthrough of the Year in 2015. [11] Many bioethical concerns have been raised about the prospect of using CRISPR for germline editing, especially in human embryos. [12]
In addition to CRISPR research, the IGI works to advance public understanding of CRISPR and genome engineering and guide the ethical use of these technologies. Free public resources include: CRISPRpedia — a free textbook-style resource for learning about the biology, applications, and ethics of CRISPR and genome editing, with chapters edited ...
In his new book “The Catalyst,” Thomas R. Cech talks about the Covid-19 vaccines, what RNA means for future health crises and how gene editing with CRISPR factors in.
Cas9 (or "CRISPR-associated protein 9") is an enzyme that uses CRISPR sequences as a guide to recognize and open up specific strands of DNA that are complementary to the CRISPR sequence. Cas9 enzymes together with CRISPR sequences form the basis of a technology known as CRISPR-Cas9 that can be used to edit genes within living organisms.
Off-target genome editing refers to nonspecific and unintended genetic modifications that can arise through the use of engineered nuclease technologies such as: clustered, regularly interspaced, short palindromic repeats ()-Cas9, transcription activator-like effector nucleases (), meganucleases, and zinc finger nucleases (ZFN). [1]
There are two main requirements for mRNA degradation to take place: a near-perfect complementary match between the guide strand and target mRNA sequence, and, a catalytically active Argonaute protein, called a 'slicer', to cleave the target mRNA. [1] There are two major pathways of mRNA degradation once cleavage has occurred.
A gRNA can and at times does have unintended interactions ("off-targets") with other locations of the genome of interest. For a given candidate gRNA, these tools report its list of potential off-targets in the genome thereby allowing the designer to evaluate its suitability prior to embarking on any experiments.