based on a bacterial CRISPR-related protein-9 nuclease (Cas9) from Streptococcus pyogenes is an effective and reliable method for making precise, targeted changes in the genome of living cells. The functions of the CRISPR (Short Palindromic Repeats Clustering Regular Intermediate Repeats) and CRISPR-associated (Cas) genes in adaptive immunity in selected bacteria and archetypes allow the organisms to respond to and remove invasive genetic material. Methods Three types of CRISPR mechanisms, the most
been found that prokaryotes, mainly bacteria and Achaea, contain an immune system that allows them to defend themselves against the nucleic acids that are integrated by viral genomes. Therefore, this discovery was made by studying the genomes of bacterial cells; which shed light on the clustered regularly interspaced short palindromic repeats (CRISPR) identified in many bacterial genomes. Therefore, the identification of CRISPR was first discovered in 1987 but its function was unknown at the time. However
Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR Associated System (CAS) is an advancement in technology that transformed a previous protein-based targeting (TALEN and Zinc Finger) process used to target and splice genes or DNA sequences. CRISPR-Cas9 was discovered in 2013 by Feng Zhang from the Broad Institute and MIT. CRISPR-Cas9 targets specific base pairs using small RNA that can be easily swapped for many different RNA targeting sites. This allowed CRISPR-Cas9 to surpass the
Clustered regularly interspaced short palindromic repeats (CRISPR) is a microbial adaptive immune system which utilizes an RNA guided Cas9 endonuclease to specifically target and destroy viral DNA via double stranded breaks (Cong et al., 2013; Hsu et al., 2014). Researchers discovered that they could manipulate the guide RNA (gRNA) to deliver Cas9 to a DNA sequence of interest within the human genome. This enables Cas9 to induce site specific double stranded breaks (Hsu et al., 2014). Donor DNA sequences