Modular custom nuclease platform for genome engineering therapies

描述(申请人提供)：基因组工程是一个新兴的领域，在这个领域中，基因组DNA序列被修改以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，然而修复过程也允许在断点处或附近对基因组序列进行受控的改变。核酸酶诱导的基因修饰目前正被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利方法，用于分离来自最近发现的一组LAGLIDADG归巢内切酶(LHE)支架的稀有切割核酸酶，与以前的LHE支架和替代技术(如锌指核酸酶和TAL效应核酸酶)相比，LHE支架具有更优越的性能(单体、高亲和力、超特异性DNA识别)，用于基因组工程治疗。这项第一阶段的SBIR重新提交提案将支持这些方法在开创性的计算模拟策略中的应用，以模拟我们全面的模块化核酸酶选择平台衍生的变体。这些模型将被用来快速构建针对基因组靶点的高度精炼的核酸酶产物。从这项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。作为计划中的第二阶段应用程序的一部分，精密基因组工程公司已经与学术计算建模实验室以及专注于干细胞和肿瘤学的实验室建立了长期合作关系，这些实验室将在大小动物多能细胞种群中进行临床前数据，从而向临床应用迈进。 与公共卫生相关：这项第一阶段SBIR提案旨在开发用于修改基因组序列的基因特异性核酸酶，这一过程被称为‘基因组工程’。基因组工程策略正在被探索用于治疗慢性病毒感染、肿瘤学应用、先天性遗传疾病的基因治疗，并且是自体干细胞治疗的一个特别有前途的工具。

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.