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序列被修饰用于生物技术和治疗目的。高度特异性核酸内切酶是实现基因组工程方法的关键工具，因为它们需要在限定的基因组序列处产生双链DNA断裂。细胞内在的DNA修复机制解决核酸酶诱导的DNA断裂，然而修复过程也能够在断点处或附近对基因组序列进行受控改变。核酸酶诱导的基因修饰目前正被探索作为慢性病毒感染的治疗、癌症免疫治疗和修复性基因治疗。Precision Genome Engineering开发了专有方法，用于分离来自最近发现的LAGLIDADG归巢核酸内切酶（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支架具有上级特性（单体、高亲和力、超特异性DNA识别），可用于基因组工程治疗。这一第一阶段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，以模拟来自我们全面的模块化核酸酶选择平台的变体。这些模型将被用来快速构建高度精细的核酸酶产品的基因组靶位点特异性。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。作为计划中的第二阶段应用的一部分，该应用将扩展本第一阶段提案中启动的工作，Precision Genome Engineering已与学术计算建模实验室以及干细胞和肿瘤学重点实验室建立了长期合作关系，这些实验室致力于在小型和大型动物多能细胞群中进行临床前数据，以实现临床应用。 公共卫生相关性：SBIR第一阶段的目标是开发用于修饰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探索慢性病毒感染的治疗，肿瘤学应用，先天性遗传疾病的基因治疗，是一个特别有前途的工具，自体干细胞治疗。

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.