RT Journal Article
SR Electronic
T1 RecV recombinase system for <em>in vivo</em> targeted optogenomic modifications of single cells or cell populations
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 553271
DO 10.1101/553271
A1 Shenqin Yao
A1 Peng Yuan
A1 Ben Ouellette
A1 Thomas Zhou
A1 Marty Mortrud
A1 Pooja Balaram
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 Andrew Curtright
A1 Ajay Dhaka
A1 Anat Kahan
A1 Viviana Gradinaru
A1 Radosław Chrapkiewicz
A1 Mark Schnitzer
A1 Hongkui Zeng
A1 Ali Cetin
YR 2019
UL http://biorxiv.org/content/early/2019/07/16/553271.abstract
AB Brain circuits are composed of vast numbers of intricately interconnected neurons with diverse molecular, anatomical and physiological properties. To allow highly specific “user-defined” 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, to create light-inducible recombinases (named RecV). 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 superficial as well as deep brain areas of live mouse brains by one-photon or two-photon light induction. These light-inducible recombinases can produce highly targeted, sparse and strong labeling of individual neurons in multiple loci and species. They can be used in combination with other genetic strategies to achieve specific intersectional targeting of mouse cortical layer 5 or inhibitory somatostatin neurons. In mouse cortex sparse light-induced recombination allows whole-brain morphological reconstructions to identify axonal projection specificity. Furthermore these enzymes allow single cell targeted genetic modifications via soma restricted two-photon light stimulation in individual cortical neurons and can be used in combination with functional optical indicators with minimal interference. In summary, RecVs enable spatiotemporally-precise, targeted optogenomic modifications that could greatly facilitate detailed analysis of neural circuits at the single cell level by linking genetic identity, morphology, connectivity and function.