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Context: Researchers at the CSIR-Institute of Genomics and Integrative Biology (CSIR-IGIB) and the L.V. Prasad Eye Institute have engineered an advanced genome-editing system.
Scientists have developed an enhanced genome-editing system that can modify DNA more precisely and more efficiently than existing CRISPR-based technologies.
CRISPR occurs naturally in some bacteria, as a part of their immune system that limits infections by recognising and destroying viral DNA.
This new system modifies DNA with greater precision and efficiency than existing CRISPR-based technologies.
CRISPR, originally part of the bacterial immune system, helps bacteria recognize and destroy viral DNA.
Scientists have repurposed this mechanism to edit genomes in higher-order organisms.
This innovation, which garnered a Nobel Prize, has significant applications in agriculture, healthcare, and more.
The CRISPR-Cas9 system, commonly used for gene editing, involves a guide RNA (gRNA) directing the Cas9 enzyme to specific DNA sequences.
The enzyme then cuts the DNA, allowing the cell’s repair system to modify the genome.
However, the widely used SpCas9 enzyme from Streptococcus pyogenes often recognizes and cuts unintended parts of the genome, leading to off-target effects.
Although scientists have developed higher fidelity versions of SpCas9, these often come at the cost of reduced editing efficiency.
Researchers have explored using the FnCas9 enzyme from Francisella novicida, which is highly precise but less efficient.
To address this, the team at CSIR-IGIB engineered new versions of FnCas9 by modifying its amino acids to enhance its binding affinity with the PAM sequence.
This modification allows the enzyme to bind more strongly to DNA, improving gene editing effectiveness.
The enhanced FnCas9, modified for greater flexibility, can access and edit harder-to-reach regions of the genome.
Experimental results showed that this version of FnCas9 cuts target DNA more effectively than the unmodified version.
It enhances the scope of CRISPR-based diagnostics and therapeutics by detecting more single-nucleotide changes in the DNA.
The enhanced FnCas9’s suitability for therapeutic use was tested by a team at the L.V. Prasad Eye Institute.
They edited the genomes of human kidney and eye cells, finding that the modified enzyme performed better than SpCas9 and exhibited negligible off-target effects.
The team also used this enhanced enzyme to correct a mutation in the RPE65 gene, which causes Leber congenital amaurosis type 2 (LCA2), an inherited form of blindness.
Researchers isolated skin cells from an individual with LCA2 and reprogrammed them into induced pluripotent stem cells (iPSCs).
These cells, differentiated into retinal cells, showed normal levels of RPE65 protein after treatment with the enhanced FnCas9.
The high efficiency and low off-target effects of the edited iPSCs indicated the potential for this technology to treat genetic disorders effectively.
The development of an enzyme with high precision and minimal off-target effects is a significant advancement for the research community.
This precision is crucial for correcting mutations in genetic diseases.
The next focus for researchers is on developing proficient delivery systems to target cells’ nuclei accurately.
The CSIR-IGIB team is adapting the system for various delivery methods and reducing the size of the enhanced FnCas9.
Collaborations with Indian companies aim to scale up and manufacture affordable therapeutic solutions for multiple genetic disorders.
This initiative, driven by indigenous intellectual property, positions India to develop cost-effective therapies for low- and middle-income countries.
By: Shubham Tiwari ProfileResourcesReport error
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