Modular custom nuclease platform for genome engineering therapies

描述（由申请人提供）：基因组工程是一个新兴领域，其中基因组DNA序列被修改用于生物技术和治疗目的。高度特异的内切酶是实现基因组工程方法的关键工具，因为它们需要在定义的基因组序列上产生双链DNA断裂。细胞内在的DNA修复机制解决了核酸酶诱导的DNA断裂，然而，修复过程也使基因组序列在断裂点或附近发生可控的改变。核酸酶诱导的基因修饰目前正在探索作为慢性病毒感染，癌症免疫治疗和修复性基因治疗的治疗方法。精密基因组工程公司开发了一种专有的方法，用于分离来自最近发现的LAGLIDADG归巢内切酶（LHE）支架的稀有切割核酸酶，与以前的LHE支架和锌指核酸酶和TAL效应核酸酶等替代技术相比，这些支架具有优越的特性（单体，高亲和力，超特异性DNA识别），可用于基因组工程治疗。这个第一阶段的SBIR重新提交提案将支持这些方法在一个开创性的计算模拟策略中的应用，以模拟来自我们的综合模块化核酸酶选择平台的变体。这些模型将被用于快速构建高度精炼的核酸酶产品，特定于基因组目标位点。从SBIR提案中产生的技术将直接转化为具有商业潜力的多种应用。精密基因组工程公司已经与学术计算建模实验室以及干细胞和肿瘤学实验室建立了长期合作关系，这些实验室将在小型和大型动物多能细胞群体中开展临床前数据研究，以推进临床应用。

DESCRIPTION (provided by applicant): Genome engineering is an emerging field in which genomic DNA sequences are modified for biotechnological and therapeutic purposes. Highly specific endonucleases are the key tool that enables genome engineering approaches, as they are required to create double-stranded DNA breaks at defined genomic sequences. Cell-intrinsic DNA repair machinery resolves nuclease-induced DNA breaks, however the repair process also enables controlled alterations to genomic sequences at or near the breakpoint. Nuclease-induced gene modifications are currently being explored as treatments for chronic viral infection, cancer immunotherapy, and reparative gene therapy. Precision Genome Engineering has developed proprietary methods for isolating rare cutting nucleases derived from a recently discovered group of LAGLIDADG homing endonuclease (LHE) scaffolds with superior properties (monomeric, high affinity, ultra-specific DNA recognition) for genome engineering therapies compared with previous LHE scaffolds and alternate technologies such as zinc-finger nucleases and TAL effector nucleases. This phase-I SBIR resubmission proposal will support the application of these methods in a pioneering computational simulation strategy to model variants derived from our comprehensive modular nuclease selection platform. These models will be exploited to rapidly construct highly refined nuclease products specific for genomic target sites. The technology emerging from this SBIR proposal will be directly translatable to multiple applications with commercial potential. As part of a planned phase-II application that would extend of work initiated in this phase-I proposal, Precision Genome Engineering has established long-term collaborations with academic computational modeling laboratories in addition to stem cell and oncology focused laboratories positioned to carry out pre-clinical data in small and large animal pluripotent cell populations to progress towards clinical applications. PUBLIC HEALTH RELEVANCE: This Phase-I SBIR proposal aims to develop gene-specific nucleases for modifying genomic sequences, a process termed 'genome engineering'. Genome engineering strategies are being explored in the treatment of chronic viral infection, oncology applications, gene therapy for inborn genetic disorders, and are an especially promising tool for autologous stem cell therapies.