Zell- und Molekularbiologie

Abstrakt

CRISPR/Cas9 Genetic Engineering Relevance to Mitigation of Human Disease and Disorder

Sanjeev Patel

The mechanism of “Clustered Regularly Interspaced Palindromic Repeat” (CRISPR) was first elucidated in the year 1987. Originally, CRISPR/Cas system is an adaptive immune molecular mechanism found to occur among 40% of bacteria and 90% of archaea that resist foreign genetic plasmids and invasion by phages. They have an. Important role in protecting the integrity of the inherent genetic constitution and factors. This structure recognizes the DNA of the phage and binds with it through CRISPR RNA also called crRNA. And then guides the CRISPR-Associated protein (Cas) to recognize and cleave the exogenous DNA by trans-activation of crRNA. There are two Cas protein families, class 1 (multi sub-unit effector complexes) and Class 2 (single protein effector modules). CRISPR/Cas9 belongs to the class 2 family that requires single guide RNA (sgRNA) to cleave the target gene. Cas9-crRNA can cut the target. DNA under in vitro conditions causing breaks that occur at three nucleotides upstream of the Protospacer Adjacent Motif (PAM). Therefore, it can edit the genes in a test tube. Later on, the efficiency of the system was improved and applied for editing the genome of eukaryotic cells, animal models for gene therapy, and plant genetic trait improvement [1]. Charpentier and Doudna were awarded Nobel Prize in Chemistry in recognition of their significant contributions to CRISPR/Cas9 based gene editing knowledge in the year 2020.