Efficient photoactivatable Dre recombinase for cell type-specific spatiotemporal control of genome engineering in the mouse

Precise genetic engineering in specific cell types within an intact organism is intriguing yet challenging, especially in a spatiotemporal manner without the interference caused by chemical inducers. Here we engineered a photoactivatable Dre recombinase based on the identification of an optimal spli...

Full description

Saved in:
Bibliographic Details
Published inProceedings of the National Academy of Sciences - PNAS Vol. 117; no. 52; pp. 33426 - 33435
Main Authors Li, Huiying, Zhang, Qiansen, Gu, Yiran, Wu, Yingyin, Wang, Yamei, Wang, Liren, Feng, Shijie, Hu, Yaqiang, Zheng, Yansen, Li, Yongmei, Ye, Haifeng, Zhou, Bin, Lin, Longnian, Liu, Mingyao, Yang, Huaiyu, Li, Dali
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 29.12.2020
Subjects
Animals
Biological Sciences
Brain - metabolism
Dependovirus - metabolism
Escherichia coli Proteins - metabolism
Gene Expression
Genes, Reporter
Genetic Engineering
Genetic Vectors - metabolism
Genome
HEK293 Cells
Humans
Integrases - metabolism
Light
Liver - metabolism
Mice, Inbred C57BL
Mice, Transgenic
Neurons - metabolism
Recombinases - metabolism
Time Factors
photoactivatable Dre recombinase
cell labeling
optogenetics
Online AccessGet full text

Cover

More Information
Summary:Precise genetic engineering in specific cell types within an intact organism is intriguing yet challenging, especially in a spatiotemporal manner without the interference caused by chemical inducers. Here we engineered a photoactivatable Dre recombinase based on the identification of an optimal split site and demonstrated that it efficiently regulated transgene expression in mouse tissues spatiotemporally upon blue light illumination. Moreover, through a double-floxed inverted open reading frame strategy, we developed a Cre-activated light-inducible Dre (CALID) system. Taking advantage of well-defined cell-type–specific promoters or a well-established Cre transgenic mouse strain, we demonstrated that the CALID system was able to activate endogenous reporter expression for either bulk or sparse labeling of CaMKIIα-positive excitatory neurons and parvalbumin interneurons in the brain. This flexible and tunable system could be a powerful tool for the dissection and modulation of developmental and genetic complexity in a wide range of biological systems.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
1H.L., Q.Z., Y.G. contributed equally to this work.
Author contributions: H.L., M.L., H. Yang, and D.L. designed research; H.L., Q.Z., Y.G., Y. Wu, Y. Wang, L.W., S.F., Y.H., Y.Z., Y.L., and H. Yang performed research; B.Z. contributed new reagents/analytic tools; H.L., Q.Z., H. Ye, B.Z., L.L., H. Yang, and D.L. analyzed data; and H.L., Q.Z., and D.L. wrote the paper.
Edited by Lei Wang, University of California San Francisco Medical Center, San Francisco, CA, and accepted by Editorial Board Member Tak W. Mak November 11, 2020 (received for review March 2, 2020)
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.2003991117