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光遗传学控制早期胚胎标记:使用光激活 Cre 重组酶 3.0。

Optogenetic control of early embryos labeling using photoactivatable Cre recombinase 3.0.

机构信息

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.

出版信息

FEBS Open Bio. 2024 Nov;14(11):1888-1898. doi: 10.1002/2211-5463.13862. Epub 2024 Sep 2.

DOI:10.1002/2211-5463.13862
PMID:39223831
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11532978/
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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04eb/11532978/0f8227718944/FEB4-14-1888-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04eb/11532978/d817686d1d4d/FEB4-14-1888-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04eb/11532978/926dbc1875a0/FEB4-14-1888-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04eb/11532978/ff5160015ce0/FEB4-14-1888-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04eb/11532978/0f8227718944/FEB4-14-1888-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04eb/11532978/d817686d1d4d/FEB4-14-1888-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04eb/11532978/926dbc1875a0/FEB4-14-1888-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04eb/11532978/ff5160015ce0/FEB4-14-1888-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/04eb/11532978/0f8227718944/FEB4-14-1888-g002.jpg

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本文引用的文献

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Optogenetics for transcriptional programming and genetic engineering.光遗传学用于转录编程和基因工程。
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Repetitive short-pulsed illumination efficiently activates photoactivatable-Cre as continuous illumination in embryonic stem cells and pre-implantation embryos of transgenic mouse.重复短脉冲光照在转基因小鼠胚胎干细胞和植入前胚胎中比连续光照更有效地激活光激活型 Cre。
Genesis. 2021 Dec;59(12):e23457. doi: 10.1002/dvg.23457. Epub 2021 Oct 23.
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Photoactivatable Cre knock-in mice for spatiotemporal control of genetic engineering in vivo.光激活 Cre 敲入小鼠用于体内遗传工程的时空控制。
Lab Invest. 2021 Jan;101(1):125-135. doi: 10.1038/s41374-020-00482-5. Epub 2020 Sep 5.
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Tamoxifen-Activated CreERT Impairs Retinal Angiogenesis Independently of Gene Deletion.他莫昔芬激活的CreERT可独立于基因缺失损害视网膜血管生成。
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Photoactivatable Cre recombinase 3.0 for in vivo mouse applications.光激活型 Cre 重组酶 3.0 用于体内小鼠应用。
Nat Commun. 2020 May 1;11(1):2141. doi: 10.1038/s41467-020-16030-0.
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Establishment of a tTA-dependent photoactivatable Cre recombinase knock-in mouse model for optogenetic genome engineering.建立一个 tTA 依赖性光激活 Cre 重组酶敲入小鼠模型,用于光遗传学基因组工程。
Biochem Biophys Res Commun. 2020 May 21;526(1):213-217. doi: 10.1016/j.bbrc.2020.03.015. Epub 2020 Mar 20.
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