Biology Of The Crispr Cas9 System - 866 Words

BIOLOGY OF THE CRISPR-CAS9 SYSTEM Functionality of the CRISPR system The endonuclease Cas9 is the core multifunctional protein of the CRISPR-Cas9 system. Two nuclease domains: HNH (39) and RuvC-like (40), within the Streptococcus pyogenes Cas9 (SpCas9) protein conduct the function of cleaving DNA strands. Studies have confirmed that the tracrRNA: crRNA duplex structure will lead to the sequence-specific DNA cleavage (41). PAM short sequence facilitates crRNA to recognize the targeted DNA sequence (42). Cas9 protein cuts the specific DNA strand that is complementary to the 20bp crRNA sequence via HNH domain; on the other hand, the SpCas9 RuvC-like domain cleaves the opposite targeted DNA strand (42) (Fig. 4). Mutating either of these two nucleases domains of SpCas9 will introduce a single-stranded DNA nick, while mutating both HNH and RuvC-like domains will transform SpCas9 into a RNA-guided DNA binding protein (42). X-ray crystallography and has confirm that endonuclease Cas9 undergoes a series of conformational rearrangements when it binds to the gu ide RNA and target DNA (55). With the development of the CRISPR-Cas9 technology, the duplex structure tracrRNA: crRNA has been engineered as one sgRNA retaining both tracrRNA and crRNA’s features. 20 nucleotides at the 5’ side of sgRNA will act as crRNA: recognizing and guiding SpCas9 protein to the targeted DNA sequences through Watson-Crick base pairing; whereas RNA forms a double-stranded structure at the 3’ end of sgRNAShow MoreRelatedDna Editing System For A Type II Crispr System Mechanism1295 Words   |  6 PagesCRISPR/Cas9 DNA editing system is a prokaryotic immune system that becomes resistant to foreign genetics such as plasmids. CRISPR stands for Clustered Regularly-Interspaced Short Palindromic Repeats, which are prokaryotic DNA strands that have a short repetition base sequence. Cas9 is a nuclease enzyme that is used to cut into DNA strands that are associated with CRISPR type II. CRISPR/Cas9 system has several advantages when using this technology in the biological and biomedi cal sciences. It alsoRead MoreAnalyzing The Genome Of Living Cells ( 1 )1139 Words   |  5 PagesBiology, in all of its glory, is quite amazing. It has always existed and always will; it merely just waits for a human to attempt to understand it. This understanding has taken centuries, however it seems to increase with the years. A very popular topic amongst biologists today is the genome, understanding it, mapping it, comparing one organism’s to another and so on. With the understanding of this genome though, we as humans want to delve into it, tweak it, and manipulate it until it is perfectionRead MoreGenes And Their Effects On Gene Editing Essay1428 Words   |  6 Pagesfields of molecular biology, as manipulated genes deem possible studying the specific genes and their effects. 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