Indian Scientists CRISPR Enzyme Discovery – Better Genetic Editing
New Delhi, India — Researchers at the L.V. Prasad Eye Institute and the CSIR-Institute of Genomics and Integrative Biology (IGIB), New Delhi, have achieved significant advances in genome-editing technology. New CRISPR enzyme variants that have been developed hold the potential to be more accurate and effective than current techniques.
The CSIR-IGIB team’s developed genome-editing technique that has the ability to change DNA more precisely and effectively. CRISPR has been repurposed to modify the genomes of higher-order species. It is a defensive system against viral infections that naturally happens in some bacteria. This technique may be used extensively in both agriculture and healthcare, allowing for the treatment of genetic abnormalities as well as advances in plant nutrition.
Resolving the Off-Target Issue with CRISPR
A guide-RNA (gRNA) is used by the CRISPR-Cas9 system, a powerful tool for introducing, eliminating, or changing DNA sequences, to target certain regions of the genome. The Cas9 enzyme is guided to the target location by the gRNA, where it executes precise cuts. But this approach frequently has off-target consequences, particularly when Streptococcus pyogenes’ SpCas9 enzyme is used. Although scientists have created SpCas9 variants with more accuracy, these advancements sometimes come at the expense of editing efficiency.
Changing to FnCas9 to Improve Precision
At CSIR-IGIB, researchers under the direction of Debojyoti Chakraborty and Souvik Maiti have concentrated on FnCas9, a kind of Cas9 enzyme from Francisella novicida that has been shown to have lesser efficiency but greater accuracy. The scientists changed the amino acids in FnCas9 that interact with the genome’s PAM region to improve the protein’s binding affinity and increase the efficiency of gene editing. Moreover, the modified FnCas9 has the ability to reach and change previously unmodifiable genomic regions.
Enhanced FnCas9 in Action
The results of experiments showed that the improved FnCas9 was able to cut target DNA more quickly than the unmodified form. The improved FnCas9 demonstrated a capacity to recognize nearly twice as many genetic alterations in diagnostic applications, expanding its range for locating mutations that cause diseases.
Potential Treatment for Hereditary Blindness
A group at the L.V. Prasad Eye Institute under the direction of Indumathi Mariappan examined the therapeutic potential of the improved FnCas9. By modifying the human kidney and eye cells’ genomes using the enzyme, the researchers were able to improve editing rates while minimizing off-target consequences. They also investigated its potential to treat Leber congenital amaurosis type 2 (LCA2), a hereditary form of blindness, by addressing a genetic abnormality. The RPE65 protein was present at normal levels in the modified cells, suggesting that the mutation was successfully corrected.
Promising Results and Future Directions
The majority of the modified cells had the required mutations with almost any off-target interactions, shocking the researchers with the improved FnCas9 enzyme’s effectiveness. According to the team’s findings, diseases like LCA2 may be treated by transplanting back person-specific retinal cells that have been altered using this same approach.
Right now, Dr. Chakraborty’s group is focusing on streamlining the delivery mechanisms and shrinking the upgraded FnCas9 system. In order to scale up and provide therapeutic solutions and offer accessible therapies for genetic diseases in low- and middle-income nations, the researchers are also in discussions with Indian businesses.
Jawaharlal Nehru University researcher Shailja Singh emphasized the significance of accuracy in CRISPR-based treatments and advocated for more attention to effective delivery methods. With the use of this indigenous invention, Indian researchers would be able to create affordable medicines, which might completely alter the way that genetic diseases are treated globally.
Indian Scientists CRISPR Enzyme Discovery – Better Genetic Editing
