Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Biomedical Synthetic Biology Research Center, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Dongchuan Road 500, Shanghai, 200241, China.
Department of Breast Surgery, Tongji Hospital, School of Medicine, Tongji University, Xincun Road 389, Shanghai, 200065, China.
Nat Commun. 2024 Nov 27;15(1):10310. doi: 10.1038/s41467-024-54781-2.
Red light optogenetic systems are in high demand for the precise control of gene expression for gene- and cell-based therapies. Here, we report a red/far-red light-inducible photoswitch (REDLIP) system based on the chimeric photosensory protein FnBphP (Fn-REDLIP) or PnBphP (Pn-REDLIP) and their interaction partner LDB3, which enables efficient dynamic regulation of gene expression with a timescale of seconds without exogenous administration of a chromophore in mammals. We use the REDLIP system to establish the REDLIP-mediated CRISPR-dCas9 (REDLIP) system, enabling optogenetic activation of endogenous target genes in mammalian cells and mice. The REDLIP system is small enough to support packaging into adeno-associated viruses (AAVs), facilitating its therapeutic application. Demonstrating its capacity to treat metabolic diseases, we show that an AAV-delivered Fn-REDLIP system achieved optogenetic control of insulin expression to effectively lower blood glucose levels in type 1 diabetes model mice and control an anti-obesity therapeutic protein (thymic stromal lymphopoietin, TSLP) to reduce body weight in obesity model mice. REDLIP is a compact and sensitive optogenetic tool for reversible and non-invasive control that can facilitate basic biological and biomedical research.
红光光遗传学系统在基因和细胞疗法的基因表达精确控制方面的需求很高。在这里，我们报告了一种基于嵌合光敏蛋白 FnBphP（Fn-REDLIP）或 PnBphP（Pn-REDLIP）及其相互作用伙伴 LDB3 的红光/远红光诱导光开关（REDLIP）系统，该系统能够在哺乳动物中无需外源给予生色团的情况下，以秒为时间尺度高效动态调控基因表达。我们使用 REDLIP 系统建立了 REDLIP 介导的 CRISPR-dCas9（REDLIP）系统，从而能够在哺乳动物细胞和小鼠中实现内源性靶基因的光遗传学激活。REDLIP 系统足够小，可以支持包装到腺相关病毒（AAV）中，从而促进其治疗应用。为了证明其治疗代谢疾病的能力，我们展示了一种 AAV 递送的 Fn-REDLIP 系统可以实现胰岛素表达的光遗传学控制，从而有效地降低 1 型糖尿病模型小鼠的血糖水平，并控制一种抗肥胖治疗性蛋白（胸腺基质淋巴细胞生成素，TSLP）来降低肥胖模型小鼠的体重。REDLIP 是一种紧凑而灵敏的光遗传学工具，可用于可逆和非侵入性的控制，从而促进基础生物学和生物医学研究。
