PhotoMorph3 and PhotoPS3 - Novel Light-Activated Antisense Agents

描述（由申请人提供）：使用靶向光来激活反义分子的基因表达的时空调节有望在透明模式生物中产生重要影响，其中照射可以在视觉上特异性地靶向已知的解剖标志。照射器官内选定的细胞，可以在生物体发育过程中的任何时间以细胞尺度分辨率关闭特定基因，从而允许对整个生物体中特定基因的作用进行深入的时空理解。这也允许研究早期发育中必要的基因，否则会导致致命的表型。我们开发了一种商用试剂、软件和仪器平台，用于进行时空基因控制实验。利用这些初步的成功，我们现在的目标是进一步开发和商业化新一代光活化反义试剂（PhotoMorph3.0和PhotoPS3.0）。所提出的光活化反义试剂是基于morpholino或硫代磷酸酯低聚物，它们的外环氢键供体和受体被保护在可破坏与目标mRNA的沃森-克里克配对的光稳定笼基上。光照射可以切割笼化基团，释放碱基参与与靶标的杂交。我们假设外环碱基保护将提供比其他光激活格式更重要的优势，包括“紧密”的关闭状态和使用自动化的方便的按需访问。第一阶段SBIR的目标是使光活化反义化学稳健地用于商业规模的合成和实施。二期SBIR的目标是为各种发育途径（例如gut400， CNS500）开发经过验证的交钥匙“工具包”，其中每种光活化反义试剂的基本表型将被描述并在公共数据库中供研究人员使用。

DESCRIPTION (provided by applicant): The spatiotemporal modulation of gene expression using targeted light to activate antisense molecules promises to have an important impact in transparent model organisms, where irradiation can be specifically targeted visually to known anatomic landmarks. Irradiating selected cells within organs permits specific genes to be turned off at any time during the development of the organism with cell-scale resolution, thereby permitting an in depth spatiotemporal understanding of the role of particular genes in the whole organism. This also allows the study of genes essential in early development that otherwise result in a lethal phenotype. We have developed a commercial reagent, software and instrumentation platform for conducting spatiotemporal gene-control experiments. Leveraging off these initial successes, we now aim to further develop and commercialize a new generation of photoactivatable antisense reagents (PhotoMorph3.0" and PhotoPS3.0"). The proposed photoactivatable antisense reagents are based on morpholino or phosphorthioate oligomers that are protected on their exocyclic hydrogen bond donors and acceptors with photolabile caging groups that disrupt Watson-Crick pairing with a target mRNA. Light irradiation cleaves the caging groups, freeing the bases to participate in hybridization with the target. We hypothesize that exocyclic base protection will offer important advantages over other photoactivatable formats, including a 'tight' off-state and facile on-demand access using automation. The goal of this Phase I SBIR is to make the photoactivatable antisense chemistry robust for commercial-scale synthesis and implementation. The goal of the Phase II SBIR will be to develop validated and turn-key "tool-kits" for various developmental pathways (e.g. gut400, CNS500), where the basic phenotype of each photoactivatable antisense reagent will be described and made available in a public database for researchers. PUBLIC HEALTH RELEVANCE: The modulation of gene expression at different times and locations using targeted light to activate molecules promises to have an important impact in transparent model organisms, where irradiation can be specifically targeted visually to known anatomic landmarks. Irradiating selected cells within organs permits specific genes to be turned off at any time during the development of the organism with cell-scale resolution, thereby permitting an in depth understanding of the role of particular genes in different locations in the whole organism and at different times during its development. We now aim to further develop and commercialize a new generation of photoactivatable chemicals for doing this (PhotoMorph3.0" and PhotoPS3.0") that we believe will offer important advantages over others.