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Cas9 (CRISPR associated protein 9, formerly called Cas5, Csn1, or Csx12) is a 160 kilodalton protein which plays a vital role in the immunological defense of certain bacteria against DNA viruses and plasmids, and is heavily utilized in genetic engineering applications.
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]
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.
Cas9 Endonuclease Dead, also known as dead Cas9 or dCas9, is a mutant form of Cas9 whose endonuclease activity is removed through point mutations in its endonuclease domains. Similar to its unmutated form, dCas9 is used in CRISPR systems along with gRNAs to target specific genes or nucleotides complementary to the gRNA with PAM sequences that ...
Targeted gene knockout using CRISPR/Cas9 requires the use of a delivery system to introduce the sgRNA and Cas9 into the cell. Although a number of different delivery systems are potentially available for CRISPR, [ 37 ] [ 38 ] genome-wide loss-of-function screens are predominantly carried out using third generation lentiviral vectors.
An RNA-guided endonuclease (e.g., Cas9 or Cas12a [13]) and its guide RNA, which can be easily altered to set the target. [5] Cas9 is the most promising technology identified in a 2014 review. [5] Cas9 gene drives have been successfully tested in 2015, [14] and Cas12a in 2023. [15]
Synthetic guide RNA is a chimera of crRNA and tracrRNA; therefore, this discovery demonstrated that the CRISPR-Cas9 technology could be used to edit the genome with relative ease. [22] Researchers worldwide have employed this method successfully to edit the DNA sequences of plants, animals, and laboratory cell lines .
On 26 November 2018, The CRISPR Journal published ahead of print an article by He, Ryan Ferrell, Chen Yuanlin, Qin Jinzhou, and Chen Yangran in which the authors justified the ethical use of CRISPR gene editing in humans. [74] As the news of CRISPR babies broke out, the editors reexamined the paper and retracted it on 28 December, announcing: