利用可光激活的 Cre 重组酶 3.0 对早期胚胎标记进行光遗传控制。

IF 2.8 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Kumi Morikawa, Akira Nagasaki, Lue Sun, Eihachiro Kawase, Tatsuhiko Ebihara, Yasuaki Shirayoshi
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引用次数: 0

摘要

建立一种高效的可光激活 Cre 重组酶 PA-Cre3.0,可实现对 Cre 重组酶活性的时空控制。这项技术有助于阐明细胞系,并促进发育过程中的基因和细胞功能分析。本研究利用 PA-Cre3.0 转基因早期小鼠植入前胚胎研究了蓝光介导的 Cre-loxP 重组光学调控。我们发现,在杂合子状态下诱导 PA-Cre3.0 表达,蓝光并不能检测到重组激活。相反,在同卵胚胎中,蓝光成功诱导了 PA-Cre3.0 的 DNA 重组,并激活了红色荧光蛋白报告基因,而在没有光照的胚胎中几乎检测不到 Cre 重组活性的泄漏。因此,我们确定了 PA-Cre3.0 系统在小鼠早期胚胎中有效发挥作用的条件。这些结果有望为某些生物学研究(如小鼠早期胚胎的发育过程分析和系谱追踪)提供一种新的光遗传学工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Optogenetic control of early embryos labeling using photoactivatable Cre recombinase 3.0

Optogenetic control of early embryos labeling using photoactivatable Cre recombinase 3.0

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.

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来源期刊
FEBS Open Bio
FEBS Open Bio BIOCHEMISTRY & MOLECULAR BIOLOGY-
CiteScore
5.10
自引率
0.00%
发文量
173
审稿时长
10 weeks
期刊介绍: FEBS Open Bio is an online-only open access journal for the rapid publication of research articles in molecular and cellular life sciences in both health and disease. The journal''s peer review process focuses on the technical soundness of papers, leaving the assessment of their impact and importance to the scientific community. FEBS Open Bio is owned by the Federation of European Biochemical Societies (FEBS), a not-for-profit organization, and is published on behalf of FEBS by FEBS Press and Wiley. Any income from the journal will be used to support scientists through fellowships, courses, travel grants, prizes and other FEBS initiatives.
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