Optogenetic Control of Transcription in Zebrafish

Abstract

Light inducible protein-protein interactions are powerful tools to manipulate biological processes. Genetically encoded light-gated proteins for controlling precise cellular behavior are a new and promising technology, called optogenetics. Here we exploited the blue light-induced transcription system in yeast and zebrafish, based on the blue light dependent interaction between two plant proteins, blue light photoreceptor Cryptochrome 2 (CRY2) and the bHLH transcription factor CIB1 (CRY-interacting bHLH 1). We demonstrate the utility of this system by inducing rapid transcription suppression and activation in zebrafish.

Algal photoreceptors: in vivo functions and potential applications

“…e Utilization of naturally occurring, light-sensitive adenylyl cyclases from the alga Euglena gracilis (euPAC) or the bacterium Beggiatoa (bPAC) to manipulate the cellular cAMP level by light (Ryu et al 2010;Schröder-Lang et al 2007;Stierl et al 2011). f Construction of a light-activated transcription system based on the (rapid) interaction of the cryptochrome CRY2 with CIB1 upon blue light irradiation (modified after Kennedy et al 2010;Liu et al 2012) of neuronal spiking (Boyden et al 2005). In combination with a light-driven bacterial chloride pump (halorhodopsin, NpHR), multiple-color optical activation and silencing of neural circuitry were achieved on the millisecond timescale in living cells and even in freely moving animals (Han and Boyden 2007;Zhang et al 2007).…”

“…In another elegant experiment, the lightdependent interaction between the blue light receptor cryptochrome and a basic helix-loop-helix transcription factor CIB1 was already used to engineer a light-induced transcription system in living mammalian cells and in zebrafish embryos (Fig. 9f) (Kennedy et al 2010;Liu et al 2012). Moreover, this approach was used for blue-light-induced protein translocation (Kennedy et al 2010) and for rapid and reversible protein oligomerization in living cells (Bugaj et al 2013).…”

TAEL: A zebrafish-optimized optogenetic gene expression system with fine spatial and temporal control

“…Several optogenetic gene expression systems have been developed (Shimizu-Sato et al, 2002;Yazawa et al, 2009;Kennedy et al, 2010;Ye et al, 2011;Ohlendorf et al, 2012;Gersbach, 2012, 2015;Liu et al, 2012;Wang et al, 2012;Chen et al, 2013;Motta-Mena et al, 2014), but so far, these systems are non-optimal for use in zebrafish. An ideal zebrafish optogenetic system would be genetically encoded, not require complicated optics or exogenous small molecules, have a large range of induction, be reversible with fairly rapid kinetics and, importantly, have little to no toxicity.…”

Reversible optogenetic control of kinase activity during differentiation and embryonic development

“…Interestingly, CRY2-CIBN binding outcompetes CRY2 oligomerization under the same light-activating conditions (Che et al, 2015). Both CRY2-CIBN heterodimerization (Boulina et al, 2013;Hughes et al, 2012;Idevall-Hagren et al, 2012;Kakumoto and Nakata, 2013;Kennedy et al, 2010;Konermann et al, 2013;Lee et al, 2014;Liu et al, 2012; and CRY2 homo-oligomerization (Bugaj et al, 2013;Chang et al, 2014;Taslimi et al, 2014;Wend et al, 2014) have been used for optogenetic control of signal transduction. It appears that CRY2-CIBN-induced protein dimerization mimics the native interaction between the two proteins better .…”

“…It appears that CRY2-CIBN-induced protein dimerization mimics the native interaction between the two proteins better . So far, the CRY2 system has been used to control transcription in Drosophila (Boulina et al, 2013), zebrafish (Liu et al, 2012) and mouse cortex (Konermann et al, 2013).…”