Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan.
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan.
FEBS Open Bio. 2024 Nov;14(11):1888-1898. doi: 10.1002/2211-5463.13862. Epub 2024 Sep 2.
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.
建立高效的光激活 Cre 重组酶 PA-Cre3.0 可以实现 Cre 重组酶活性的时空控制。这项技术可能有助于阐明细胞谱系,并在发育过程中促进基因和细胞功能分析。本研究通过转 PA-Cre3.0 早期小鼠胚胎研究了蓝光介导的 Cre-loxP 重组的光调控。我们发现,杂合状态下诱导 PA-Cre3.0 表达,不会因蓝光而检测到可检测的重组激活。相反,在纯合胚胎中,蓝光成功诱导 PA-Cre3.0 的 DNA 重组,导致红色荧光蛋白报告基因的激活,而在没有光照的胚胎中几乎检测不到 Cre 重组酶活性的泄漏。因此,我们对 PA-Cre3.0 系统在早期小鼠胚胎中高效发挥作用的条件进行了表征。这些结果有望为某些生物学研究提供新的光遗传学工具,如早期小鼠胚胎发育过程分析和谱系追踪。