Optogenetic control of early embryos labeling using photoactivatable Cre recombinase 3.0

Author:

Morikawa Kumi1ORCID,Nagasaki Akira2ORCID,Sun Lue3,Kawase Eihachiro4ORCID,Ebihara Tatsuhiko2,Shirayoshi Yasuaki5

Affiliation:

1. Cellular and Molecular Biotechnology Research Institute National Institute of Advanced Industrial Science and Technology (AIST) Tsukuba Japan

2. Biomedical Research Institute National Institute of Advanced Industrial Science and Technology (AIST) Tsukuba Japan

3. Health and Medical Research Institute National Institute of Advanced Industrial Science and Technology (AIST) Tsukuba Japan

4. Institute for Life and Medical Sciences Kyoto University Kyoto Japan

5. Division of Regenerative Medicine and Therapeutics, Department of Genomic Medicine and Regenerative Therapy, Faculty of Medicine Tottori University Yonago Japan

Abstract

Establishing a highly efficient photoactivatable Cre recombinase PA‐Cre3.0 can allow spatiotemporal control of Cre recombinase activity. This technique may help to elucidate cell lineages, as well as facilitate gene and cell function analysis during development. This study examined the blue light‐mediated optical regulation of Cre‐loxP recombination using PA‐Cre3.0 transgenic early mouse pre‐implantation embryos. We found that inducing PA‐Cre3.0 expression in the heterozygous state did not show detectable recombination activation with blue light. Conversely, in homozygous embryos, DNA recombination by PA‐Cre3.0 was successfully induced by blue light and resulted in the activation of the red fluorescent protein reporter gene, while almost no leaks of Cre recombination activity were detected in embryos without light illumination. Thus, we characterize the conditions under which the PA‐Cre3.0 system functions efficiently in early mouse embryos. These results are expected to provide a new optogenetic tool for certain biological studies, such as developmental process analysis and lineage tracing in early mouse embryos.

Publisher

Wiley

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