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

这个项目将展示超声波产生的微泡帮助视网膜细胞进行基因编辑的能力。其目标是恢复盲人视网膜组织的光敏感性，作为治疗失明的一步。这项拟议的技术有可能规避在利用最近尝试的其他基因转移方法时出现的安全担忧。蛋白质、DNA、聚合物等在微泡内的转移发生在超声场作用下的气泡破裂过程中。这导致了受损视网膜细胞的修复。迫切需要一种非竞争工具，通过一种侵入性较小的途径将治疗性基因有效地输送到眼睛中。该项目将结合微泡和超声波，以促进将Crispr/Cas9工具输送到视网膜细胞中，用于治疗眼病。他们将a：通过修改微泡和声学参数来优化RPE65 Crispr/Cas9的体外效率；b：研究功能化微泡的靶向传递，并检测这些气泡在辐射压力下穿过神经视网膜的能力；以及c：通过测试RPE65缺陷Rd12小鼠光感受器的光敏恢复来评估RPE65 Crispr/Cas9的治疗潜力。该奖项反映了NSF的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.