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treatment part 2

Submitted by ziweiwang on Thu, 12/12/2019 - 21:25

The result indicates that because the experiment was done in vivo, the results of the experiment suggest a high chance that the method would work in an organism as long as the two organisms are similar. In the third experiment that was done the ONL thickness in electroporated eyes and the non electroporated eyes were assessed through histological analysis. The result of this experiment indicated that the retina is able to undergo phototransduction. However, the researchers also observed that there was damage to the retina from the electroporation treatment. The results suggest that the treatment works in restoring vision, However, electroporation is not a viable method for use on humans in rescuing vision. The fourth experiment tested the efficiency of using AAV mediated delivery by injecting the virus with therapy that was used in previous experiments and indel frequency and TIDE analysis was done to evaluate the efficiency of this delivery method. Using this delivery method, all retinal cell types were targeted but to a differing extent. The P23H mutation had a moderate level of cleavage while wt was unaffected the P23H also had a 5 times higher amount of indel compared to wt. This indicates that the viral delivery of the treatment was as effective as electroporation and usable for human trials. In the fifth experiment that was done the NGS analysis of the edited retina was performed through a sequencing analysis. The result is that plus one and plus two insertions was the most common mutation which caused a deletion codon, indicating that it would be unlikely that there would be an inframe mutation. This indicates that the treatment method is highly selective and is effective at the silencing of the mutated protein.

 In the sixth experiment, Human cells were treated with the therapy to induce mutations into the cells using sgRNA-hWT and hMUT. The results show that both guides showed different cleavage efficiency in different analysis and there was a high specificity in targeting the complementary genes.  In addition, because most of the indel inframe mutation was very low there was a higher chance that the mutation would have no effect on the gene. Because of this result, there is a high chance that this method of delivery would also work well in the human retina. This paper allows the field of disease research to move forward by showing a method of treatment that has worked in the human cells and mice cells this would mean that this treatment system and the delivery combination is likely to work in the human eye and that the treatment can be moved into clinical trials. 

While retinitis pigmentosa has unique genetics, protein structure, and tissue behavior that result in unique challenges for both treatment providers and patients, there have also been advances that are developing to solve the problem of the illness. However, there will be a long time until a cure is found. Because of this, funding and continuation of research will be crucial.