A non-invasive far-red light-induced split-Cre recombinase system for controllable genome engineering in mice
A non-invasive far-red light-induced split-Cre recombinase system for controllable genome engineering in mice
复制标题
用于小鼠可控基因组工程的非侵入性远红光诱导分裂 Cre 重组酶系统
DOI：
10.1038/s41467-020-17530-9
复制
发表时间：
2020-07-24
影响因子：
16.6
通讯作者：
Ye, Haifeng
中科院分区：
文献类型：
--
作者：
Wu, Jiali;Wang, Meiyan;Ye, Haifeng
Cre-loxP重组系统是基因操作的有力工具。然而，化学诱导的Cre-loxP系统存在广泛认识到的局限性，并且UV和蓝光诱导的系统具有光毒性和最小的深度组织渗透能力。在这里，我们开发了一个远红光诱导的分裂Cre-loxP系统（FISC系统）的基础上的细菌光敏色素光遗传学系统和分裂Cre重组酶，使光遗传学调控的基因组工程在体内只利用远红光（FRL）。FISC系统表现出低背景和不可检测的光细胞毒性，同时提供有效的FRL诱导的DNA重组。我们的体内研究显示了FISC系统强大的器官穿透能力，在肝脏重组诱导方面明显优于两种基于蓝光的Cre系统。证明其强大的临床相关性，我们成功地部署了FISC系统使用腺相关病毒（AAV）交付。因此，FISC系统扩展了用于DNA重组的光遗传学工具箱，以在生命系统中实现时空控制的非侵入性基因组工程。目前的光诱导Cre-loxP系统具有最小的深度组织穿透能力。在这里，作者提出了一个远红光诱导的分裂Cre-loxP系统在体内基因组工程。
The Cre-loxP recombination system is a powerful tool for genetic manipulation. However, there are widely recognized limitations with chemically inducible Cre-loxP systems, and the UV and blue-light induced systems have phototoxicity and minimal capacity for deep tissue penetration. Here, we develop a far-red light-induced split Cre-loxP system (FISC system) based on a bacteriophytochrome optogenetic system and split-Cre recombinase, enabling optogenetical regulation of genome engineering in vivo solely by utilizing a far-red light (FRL). The FISC system exhibits low background and no detectable photocytotoxicity, while offering efficient FRL-induced DNA recombination. Our in vivo studies showcase the strong organ-penetration capacity of FISC system, markedly outperforming two blue-light-based Cre systems for recombination induction in the liver. Demonstrating its strong clinical relevance, we successfully deploy a FISC system using adeno-associated virus (AAV) delivery. Thus, the FISC system expands the optogenetic toolbox for DNA recombination to achieve spatiotemporally controlled, non-invasive genome engineering in living systems. Current light-inducible Cre-loxP systems have minimal capacity for deep tissue penetration. Here, the authors present a far-red light-induced split Cre-loxP system for in vivo genome engineering.