How has CRISPR-Cas9 treatment opened new avenues in

both the progress and complications of editing the

mutation causing Leber's congenital amaurosis 10?

INTRODUCTION
Scientists have been able to use genetic technologies to 'cut and paste' DNA for decades. However, the newest technology, CRISPR Cas9 aims to revolutionise treatments of
cancers and rare genetic diseases such as Leber's Congenital Amaurosis 10 (LCA10, which is the most common cause of inherited childhood blindness yet its incidences reach
only 1/100,000 births worldwide. This powerful new tool; CRISPR Cas9 has opened new avenues in genomic editing as it can delete the mutated CEP290 gene, causing the
vision loss that characterises LCA10, and add the corrected version with more precision than ever before. Although its only use in clinical trials for LCA10 suggests the
uncertainties around CRISPR remain high. Hence, this article seeks to evaluate the progress and complications CRISPR Cas9 has in the treatment of LCA10.

DISEASE PROFILE
Leber congenital amaurosis (LCA10) is a genetic eye disease that primarily affects the retina. People with this disorder typically have severe visual impairment
beginning in infancy as their eyes can no longer perceive light. The biological cause of this disease is the nonsense mutation of the CEP290 gene which acts as a stop
signal, halting the production of the protein necessary for the light-sensing cells photoreceptor cells in the retina, hence causing vision loss. Additionally, LCA10 has
autosomal recessive inheritance, hence, if both parents are carriers or possess the disease, it is possible for their offspring to have LCA10. This allows the disease to
be prevented through genetic testing before pregnancy which identifies whether the embryo acquired the mutated gene. Management protocol for affected
individuals includes procedure to correct refractive error or use of vision aids when possible. However, LCA10 is currently treated using gene therapy but the most
recent technology, CRISPR Cas9 is a treatment involving the cutting and replacing of the mutated gene with a functional gene, allowing the production of the protein
crucial to the photoreceptor.

TREND 1
USE OF TECHNOLOGY
Diagram 1.2: Flowchart of the process of CRISPR-Cas9 used in LCA10

TREND 2
 USE OF TECHNOLOGY
In reference to diagram 1.2 the mutated gene is cut out by the Cas9 nuclease
and CRISPR-Cas9 also acquires the correct CEP290 gene from a donor, and is
inserted back into the LCA10 patient. As a result, the patient now possesses
the corrected CEP290 gene with its intended function.

The CRISPR Cas9 technology effectively treats the LCA10 disease as the
repaired corrected CEP290 gene can now be transcribed into the normal
mRNA and thus is further translated into the fully functional CEP290 protein
required in the structure of photoreceptor cells. Diagram 1.3 shows how the
protein formed from the expression of the normal CEP290 forms the
connecting cilium in the photoreceptor cell. Whereas, the previously mutant
CEP290's stop signal didn't allow a full connecting cilium protein, and hence, a
dysfunctional photoreceptor.

DISCUSSION OF TECHNOLOGY
MA

CONCLUSION
Hence, this inquiry observed the ability of CRISPR Cas9 in treating Leber's Congenital Amaurosis through its ability to accurately cut and replace the mutated
CEP290 gene to produce the structural protein required to restore vision. CRISPR-Cas9 has been beneficial through its ability to improve the ease, efficiency and
genetically personalised nature of treatment, regardless of its complications surrounding fears of off-target effects and fostering income inequality. CRISPR Cas9
has proved to be a powerful tool in treating multiple patients with LCA10 from the Editas clinical trial which has initialised the first in-vivo treatment. Through
further scientific discovery CRISPR-Cas9 will experience improvements in precision and safety; where in the future researchers aim for simultaneous editing. A
Cas12a biomolecule is to be created which will enable CRISPR to edit many genes in a short space of time and thus improve the gap between the expression of
the corrected CEP290 gene in comparison to non-affected individuals' (wild type) expression of this gene, hence, advocating for its widespread use to treat
LCA10.