RT Journal Article
SR Electronic
T1 RecV recombinase system for spatiotemporally controlled light-inducible genomic modifications
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 553271
DO 10.1101/553271
A1 Ali Cetin
A1 Shenqin Yao
A1 Ben Ouellette
A1 Pooja Balaram
A1 Thomas Zhou
A1 Marty Mortrud
A1 Soumya Chatterjee
A1 Yun Wang
A1 Tanya L. Daigle
A1 Bosiljka Tasic
A1 Xiuli Kuang
A1 Hui Gong
A1 Qingming Luo
A1 Shaoqun Zeng
A1 Anat Kahan
A1 Viviana Gradinaru
A1 Hongkui Zeng
YR 2019
UL http://biorxiv.org/content/early/2019/02/18/553271.abstract
AB Brain circuits are composed of vast numbers of intricately interconnected neurons with diverse molecular, anatomical and physiological properties. To allow highly specific targeting of individual neurons for structural and functional studies, we modified three site-specific DNA recombinases, Cre, Dre and Flp, by combining them with a fungal light-inducible protein, Vivid, so that their recombinase activities can be driven by blue light. We generated viral vectors to express these light-inducible recombinases and demonstrated that they can induce genomic modifications in dense or sparse populations of neurons in live mouse brains controlled by one-photon or two-photon light induction. As an important application, we showed that light-inducible recombinases can produce highly targeted, sparse and strong labeling of individual neurons thereby enabling whole-brain morphological reconstruction to identify their axonal projection specificity. In addition to targeting cortical brain areas, we applied the method in deep targets, with a demonstration of functional calcium imaging. These molecular tools enable spatiotemporally-precise, targeted genomic modifications that will greatly facilitate detailed analysis of neural circuits and linking genetic identity, morphology, connectivity and function.