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XB-ART-52569
Development 2016 Nov 01;14321:4085-4094. doi: 10.1242/dev.140889.
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Reversible optogenetic control of kinase activity during differentiation and embryonic development.

Krishnamurthy VV , Khamo JS , Mei W , Turgeon AJ , Ashraf HM , Mondal P , Patel DB , Risner N , Cho EE , Yang J , Zhang K .


Abstract
A limited number of signaling pathways are repeatedly used to regulate a wide variety of processes during development and differentiation. The lack of tools to manipulate signaling pathways dynamically in space and time has been a major technical challenge for biologists. Optogenetic techniques, which utilize light to control protein functions in a reversible fashion, hold promise for modulating intracellular signaling networks with high spatial and temporal resolution. Applications of optogenetics in multicellular organisms, however, have not been widely reported. Here, we create an optimized bicistronic optogenetic system using Arabidopsis thaliana cryptochrome 2 (CRY2) protein and the N-terminal domain of cryptochrome-interacting basic-helix-loop-helix (CIBN). In a proof-of-principle study, we develop an optogenetic Raf kinase that allows reversible light-controlled activation of the Raf/MEK/ERK signaling cascade. In PC12 cells, this system significantly improves light-induced cell differentiation compared with co-transfection. When applied to Xenopus embryos, this system enables blue light-dependent reversible Raf activation at any desired developmental stage in specific cell lineages. Our system offers a powerful optogenetic tool suitable for manipulation of signaling pathways with high spatial and temporal resolution in a wide range of experimental settings.

PubMed ID: 27697903
PMC ID: PMC5117147
Article link: Development
Grant support: [+]

Species referenced: Xenopus
Genes referenced: cry2 mapk1 odc1 raf1 tbxt


Article Images: [+] show captions
References [+] :
Beyer, Red Light-Regulated Reversible Nuclear Localization of Proteins in Mammalian Cells and Zebrafish. 2015, Pubmed