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

这项小企业创新研究（SBIR）二期项目的更广泛影响/商业潜力将是利用一种新颖的专有精确基因编辑技术开发脊髓性肌萎缩症的治疗方法。该技术将通过优化基因编辑的精度来纠正疾病，从而提高基因编辑的安全性和有效性。这项技术可用于改善甚至治愈遗传病患者。这个小企业创新研究（SBIR）二期项目将使用一种新颖的专有CRISPR-Cas精确基因编辑系统开发和验证脊髓性肌萎缩症（SMA）的治疗方法。使用CRISPR-Cas基因编辑器操纵人类基因组已在早期临床试验中得到验证，以纠正遗传疾病。然而，这项技术的应用受到高度非预期的“脱靶”编辑事件的限制。这个问题对于体内治疗来说尤其严重，因为编辑系统将被系统地或直接传递到目标器官，从而消除了筛选细胞以获得不想要的编辑结果的能力。为了保证体内基因编辑疗法的安全性和有效性，必须开发高精度的基因编辑技术。该项目将开发一种工程抗crispr （ErAcr）蛋白，以消除意外的脱靶编辑。ErAcr蛋白将与一种新型CRISPR-Cas核酸酶配对，并开发成一种治疗SMA的疗法。该疗法将被包装成一种腺相关病毒载体，并在患者来源的细胞和SMA小鼠模型中进行编辑和SMA疾病纠正测试。有效的编辑将导致患者细胞和小鼠的疾病纠正，而不会出现可观察到的脱靶编辑事件。该奖项反映了美国国家科学基金会的法定使命，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。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project will be the development of a curative therapy for spinal muscular atrophy using a novel and proprietary precision gene editing technology. The proposed technology will increase the safety and efficacy of gene editing by optimizing the precision of editing to correct the disease. This technology can be used to improve and even cure patients of genetic disease.This Small Business Innovation Research (SBIR) Phase II project will develop and validate a curative therapy for spinal muscular atrophy (SMA) using a novel and proprietary CRISPR-Cas precision gene editing system. Manipulation of the human genome using CRISPR-Cas gene editors has been validated in early clinical trials to correct genetic diseases. However, application of this technology has been limited by the high degree of unintended 'off-target' editing events. This problem is particularly acute for in vivo therapies where the editing system will be delivered systemically or directly to the target organs, eliminating the ability to screen cells for unwanted editing outcomes. High precision gene editing technology must be developed to ensure the safety and efficacy of in vivo gene editing therapies. This project will develop an engineered anti-CRISPR (ErAcr) protein to eliminate unintended off-target editing. The ErAcr protein will be paired with a novel CRISPR-Cas nuclease and developed into a therapy for SMA The therapy will be packaged into an adeno-associated viral vector and tested for editing and SMA disease correction in patient-derived cells and in an SMA mouse model. Effective editing will lead to disease correction in both patient cells and mice with no observable off-target editing events.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.