SBIR Phase I: Development of a safe gene editing system via CRISPR-Cas and Cas inhibitor co-delivery

SBIR 第一阶段：通过 CRISPR-Cas 和 Cas 抑制剂共同传递开发安全的基因编辑系统

• 批准号: 2015148
• 负责人: David Rabuka
• 金额: $ 22.5万
• 依托单位: ACRIGEN BIOSCIENCES, INC.
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-15 至 2021-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2015148&HistoricalAwards=false
• 关键词: SBIR; Phase; Development; safe; gene

The broader impact of this Small Business Innovation Research (SBIR) Phase I project is to enable safe and effective human gene editing, providing therapies targeting myriad human genetic diseases. CRISPR (clustered, regularly interspaced, short palindromic repeats) and CRISPR-associated (Cas) genes compose adaptive microbial `immune systems' found in diverse bacterial species, serving as a defense mechanism against viral infection. The simplicity, programmability, and versatility of CRISPR-Cas systems have enabled genetic modification of many organisms and offer immense therapeutic potential for treating human diseases. However, CRISPR-based gene editing can also cause off-target edits, resulting in the introduction of mutations, insertions, deletions, or DNA restructuring at unintended off-target editing. This effect can cause significant problems. The proposed technology will develop tools to safely translate CRISPR-based gene editing to in vivo human therapeutics. This Small Business Innovation Research (SBIR) Phase I project is to advance a technology based on virus-encoded CRISPR-Cas inhibitor off-switches, enabling control of off-target gene editing. These anti-CRISPR proteins are a novel class of robust protein inhibitors that can be genetically encoded for co-delivery with the CRISPR editing machinery. This proposal will focus on understanding CRISPR editing kinetics to pinpoint the ideal ‘editing window’ providing the highest therapeutic benefit while minimizing off-target risk. Second, the project will design a system for simultaneous delivery of the CRISPR editing machinery along with the anti-CRISPR inhibitor to leverage this new ‘editing window’. Genetic regulatory elements will be used to tune the expression of the various components of the system and provide the framework for a therapeutic delivery system. This work will enable safe and effective gene editing in a therapeutically relevant context, providing a toolkit for translation.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 criteria.

小企业创新研究（SBIR）第一阶段项目的更广泛影响是实现安全有效的人类基因编辑，提供针对多种人类遗传疾病的治疗方法。CRISPR（成簇的、规则间隔的、短回文重复序列）和CRISPR相关（Cas）基因组成了在不同细菌物种中发现的适应性微生物“免疫系统”，作为抵抗病毒感染的防御机制。CRISPR-Cas系统的简单性、可编程性和多功能性使许多生物体能够进行遗传修饰，并为治疗人类疾病提供了巨大的治疗潜力。然而，基于CRISPR的基因编辑也可能导致脱靶编辑，导致在非预期的脱靶编辑中引入突变、插入、缺失或DNA重组。 这种影响可能会导致严重的问题。该技术将开发工具，将基于CRISPR的基因编辑安全地转化为体内人类治疗方法。这项小型企业创新研究（SBIR）第一阶段项目旨在推进一项基于病毒编码的CRISPR-Cas抑制剂关闭开关的技术，从而能够控制脱靶基因编辑。这些抗CRISPR蛋白是一类新型的稳健的蛋白质抑制剂，其可以被遗传编码用于与CRISPR编辑机制共递送。该提案将专注于了解CRISPR编辑动力学，以确定理想的“编辑窗口”，提供最高的治疗益处，同时最大限度地减少脱靶风险。其次，该项目将设计一个系统，用于同时提供CRISPR编辑机制沿着抗CRISPR抑制剂，以利用这个新的“编辑窗口”。遗传调控元件将用于调节系统的各种组分的表达，并为治疗递送系统提供框架。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。