CRISPR/CAS9 in Toxoplasma gondii - Article Example

CRISPR- Cas 9 system is used for the production of recombinant proteins in Humans, Mice, Caenorhabditis elegans, Saccharomyces cerevisiae, Zebra Fish, Drosophila melanogaster, Sorghum, Wheat and Arabidopsis. The efficiency and the precision of genome editing by CRISPR/Cas 9 system in the tested organisms are very high. Thus, the genome editing of Toxoplasma Gondii using CRISPR/Cas9 system can be performed. CRISPR, which stands for Clustered Regularly Interspaced Short Palindromic Repeats, was identified around 1980s in the bacteria.

(Barrangou & Oost, 2012). CRISPR helps to turn on the active components, which are present in between the spacer DNA, as repeats. The CRISPR fragments get easily incorporated into the plasmids, viruses and other genomes. These spacer elements can be used to create small silencing RNA molecules (siRNAs) to prevent virus replication. Similarly, the bacteria can be made resistant to virus by introducing these small spacer elements. Thus, CRISPR plays an important role in the functioning of the immune system, and it is generally described as genome immune system.

(Shapiro, 2011). CRISPR spacers get associated with many Cas proteins and acts as auto immune system in the body. Six core Cas proteins are identified so far and the total Cas family count is expected to be up to 45. In a microbial genome, there will be more than one CRISPR- Cas locus. Cas 1 and Cas 2 proteins are considered as the important markers for CRISPR loci, with Cas 1 being the most conserved Cas protein and is present in all CRISPR loci. (Gardner & Olson, 2012). CAS 1 and Cas 2 are metal dependent as well as DNA specific, and so are highly conserved endonucleases.

They require either manganese or magnesium ion to function. In addition, Cas 1 and CAS 2 require one more Cas protein to function. As far as other proteins are concerned, Cas3 protein contains ATP- dependent helicase and Mg- dependent ssDNA nuclease. (Gardner & Olson,