CRISPR Cas9: Science & Beyond, The CRISPR cas9 also pronounced as “crisper” system is a tool for cutting DNAs in a targeted place. However, scientists always look for new opportunities, so what else CRISPR can do? since the CRISPR Cas9 bacteria immune system has already been found to be adapted to a powerful genome-research tool.
There are two components to the system a DNA cutting protein called Cas9 and an RNA molecule known as the guide RNA bound together they form a complex that can identify and cut specific sections of DNA.
Cas9 must first identify and bind to a common sequence known as PAM in the genome. Once the RNA guides are bound to the RNA guide, it unwinds part of the double helix, and the RNA strand is designed to match and bind a sequence in the DNA, once it finds the correct sequence of the DNA, Cast9 can cut into the nuclear areas.
The CRISPR-Cas9 mechanisms recognize the DNA targets that complement the short CRISPR RNA (crRNA) sequences. The complementary part of the crRNA sequence of the target sequence is called a spacer. For Cas9 to work, a specific adjacent motive for the protospacer adjacent motif (PAM) is required, which varies depending on the gene’s bacterial species.
The process of remediation is error susceptible, often introduce mutations which inadvertently disable the gene making CRISPR a fine tool to kill particular genes, but making double strand breaks is a fact that all CRISPRs can do to turn off the cas9 domains, or to fuse new enzymes into protein cast.
In a single example, Cas9 is fused into a deaminase enzyme which mutates specific DNA bases that will eventually replace adine with fibre dean, which means that you can turn a disease-related mutation into a healthy version of the gene, or put a stop codon at a certain location.
The activators can be recruited instead as guides of RNA either by fusing the cell’s transcription machine to achieve RNA polymerase and furthermore through a series of peptides. Instead, the same principle applies to gene silencing by adding transcript activators to cas9 through fusing it directly or by means of a string of peptide.
A more out-of-box idea for using CRISPR is to attach fluorescent proteins to the complex so that you can look where specific DNA sequences can be found in the cell, which can help you visualize the genome’s 3d-architecture or paint a whole chromosome, and to follow the position of the CRISPR core.
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