Microbubble-assisted Crispr/Cas9 Delivery in Retina for Photoreceptor Therapeutics

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This project will demonstrate the ability of microbubbles generated by ultrasound to aid in gene editing of retinal cells. The goal is to restore light sensitivity to blind retina tissues, as a step to toward curing blindness. The proposed technology has the potential to circumvent safety concerns that arise when utilizing other gene transport approaches that were recently attempted. The transfer of proteins, DNA, polymer etc., within the microbubbles occurs during the collapse of the bubble under the ultrasound field. This leads to repair of damaged retinal cells.There is a dire need of a nonrival vehicle to efficiently transport therapeutic genes into the eye via a less invasive route. The project will combine microbubbles and ultrasound to facilitate the delivery of Crispr/Cas9 tools into retinal cells for the treatment of ocular diseases. They will a: optimize the efficiency of RPE65 Crispr/Cas9 in vitro by modifications of microbubbles and acoustic parameters, b: investigate targeted delivery with functionalized microbubbles and examine the ability of the bubbles to pass through neural retina under radiation force, and c: evaluate the therapeutic potential of RPE65 Crispr/Cas9 by testing light sensitivity restoration of photoreceptors in RPE65 deficient rd12 mice. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteriaThis award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.