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

微泡辅助 Crispr/Cas9 在视网膜中的递送用于光感受器治疗

• 批准号: 1805210
• 负责人: Mark Humayun
• 金额: $ 48万
• 依托单位: University of Southern California
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1805210&HistoricalAwards=false
• 关键词: Microbubble; assisted; Crispr; Cas9; Delivery

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.

该项目将展示超声波产生的微泡帮助视网膜细胞基因编辑的能力。目标是恢复失明视网膜组织的光敏感性，作为治愈失明的一步。 所提出的技术有可能规避在利用最近尝试的其他基因转运方法时出现的安全问题。蛋白质、DNA、聚合物等的转移，在超声波场下气泡的破裂过程中发生。这导致受损视网膜细胞的修复，迫切需要一种非竞争性的载体，通过侵入性较小的途径将治疗基因有效地输送到眼睛中。该项目将联合收割机微泡和超声波相结合，促进Crispr/Cas9工具进入视网膜细胞，用于治疗眼部疾病。他们将：通过修饰微泡和声学参数优化体外RPE 65 Crispr/Cas9的效率，B：研究用功能化微泡的靶向递送并检查在辐射力下气泡穿过神经视网膜的能力，以及c：通过测试RPE 65缺陷型rd 12小鼠中光感受器的光敏感性恢复来评估RPE 65 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知识价值和更广泛的影响审查标准进行评估，被认为值得支持该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。