TY - JOUR
T1 - RecV recombinase system for in vivo targeted optogenomic modifications of single cells or cell populations
AU - Yao, Shenqin
AU - Yuan, Peng
AU - Ouellette, Ben
AU - Zhou, Thomas
AU - Mortrud, Marty
AU - Balaram, Pooja
AU - Chatterjee, Soumya
AU - Wang, Yun
AU - Daigle, Tanya L.
AU - Tasic, Bosiljka
AU - Kuang, Xiuli
AU - Gong, Hui
AU - Luo, Qingming
AU - Zeng, Shaoqun
AU - Curtright, Andrew
AU - Dhaka, Ajay
AU - Kahan, Anat
AU - Gradinaru, Viviana
AU - Chrapkiewicz, Radosław
AU - Schnitzer, Mark
AU - Zeng, Hongkui
AU - Cetin, Ali
N1 - Publisher Copyright:
© 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2020/4/1
Y1 - 2020/4/1
N2 - Brain circuits comprise vast numbers of interconnected neurons with diverse molecular, anatomical and physiological properties. To allow targeting of individual neurons for structural and functional studies, we created light-inducible site-specific DNA recombinases based on Cre, Dre and Flp (RecVs). RecVs can induce genomic modifications by one-photon or two-photon light induction in vivo. They can produce targeted, sparse and strong labeling of individual neurons by modifying multiple loci within mouse and zebrafish genomes. In combination with other genetic strategies, they allow intersectional targeting of different neuronal classes. In the mouse cortex they enable sparse labeling and whole-brain morphological reconstructions of individual neurons. Furthermore, these enzymes allow single-cell two-photon targeted genetic modifications and can be used in combination with functional optical indicators with minimal interference. In summary, RecVs enable spatiotemporally precise optogenomic modifications that can facilitate detailed single-cell analysis of neural circuits by linking genetic identity, morphology, connectivity and function.
AB - Brain circuits comprise vast numbers of interconnected neurons with diverse molecular, anatomical and physiological properties. To allow targeting of individual neurons for structural and functional studies, we created light-inducible site-specific DNA recombinases based on Cre, Dre and Flp (RecVs). RecVs can induce genomic modifications by one-photon or two-photon light induction in vivo. They can produce targeted, sparse and strong labeling of individual neurons by modifying multiple loci within mouse and zebrafish genomes. In combination with other genetic strategies, they allow intersectional targeting of different neuronal classes. In the mouse cortex they enable sparse labeling and whole-brain morphological reconstructions of individual neurons. Furthermore, these enzymes allow single-cell two-photon targeted genetic modifications and can be used in combination with functional optical indicators with minimal interference. In summary, RecVs enable spatiotemporally precise optogenomic modifications that can facilitate detailed single-cell analysis of neural circuits by linking genetic identity, morphology, connectivity and function.
UR - http://www.scopus.com/inward/record.url?scp=85083042623&partnerID=8YFLogxK
U2 - 10.1038/s41592-020-0774-3
DO - 10.1038/s41592-020-0774-3
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C2 - 32203389
AN - SCOPUS:85083042623
SN - 1548-7091
VL - 17
SP - 422
EP - 429
JO - Nature Methods
JF - Nature Methods
IS - 4
ER -