Actiflash

The stable Tamoxifen-like photoactivable inducer to perform a spatial and temporal control of your favorite proteins under illumination

Overview

Odoo • A picture with a caption

Actiflash is a tamoxifen-like caged-inducer, called cyclofen, that enables, upon photoactivation, the fast releasing of proteins fused to the modified estrogen receptor ligand-binding domains, ERT2. Optical-induced uncaging of Actiflash releases the fusion proteins, allowing them to translocate into the nucleus, activate transcription (using Gal4-UAS system) or induce recombination (using the Cre-lox one).

The protein expression can thus be simply controlled in time and space thanks to UV-light. 

Actiflash is a great R&D tool to convert your inducible ERT model into a photo-inducible one. It is very helpful if you want to control transcription (using Gal4-UAS) or induce recombination (using Cre-lox) in space and/or time for in-vivo cell tracking experiments and more.    

Visit our  glossary  for a definition of Tamoxifen.

A technology designed by   Ludovic Jullien Isabelle Aujard  and  Thomas Le Saux
Published in 
ChemBioChem / Molecules

Images

Features


Wide applicative scope


Technology capitalizing on the versatile use of Tamoxifen-OH for controlling functions of multiple types of proteins.


Simple conditioning


Caged Cyclofen-OH is cell-permeant and can be added either in the external medium or directly injected for conditioning.


Excellent chemical stability

Caged Cyclofen-OH does not generate any basal activation of protein function and it benefits from an excellent temporal resolution upon uncaging.


Favorable wavelength ranges for uncaging

Uncaging requires either UV-A light or a strong IR laser. Visible light is inactive, which facilitates the experiments with biological samples.

Photochemical stability

Caged Cyclofen-OH liberates Cyclofen-OH, which is photostable in contrast to Tamoxifen-OH encountering photodegradation under illumination.

Protocol

Calibration of the Actiflash concentration
It is advised to first establish the extent of phenotype sought for as a function of the Tamoxifen-OH concentration. Then the concentration of Actiflash used for sample conditioning is fixed at Tamoxifen-OH concentration causing 100% of the desired phenotype (in general 3-5 μM in cultured cells and zebrafish embryos).

Conditioning protocol with Actiflash
Incubate your samples in a serum-free medium for 90 mins, away from light.

Actiflash photoactivation
Illumination of Actiflash may be performed with UV (325-425nm range) light or multiphoton excitation (at 750 and 1064 nm with two- and three-photon excitation, respectively) to release Cyclofen-OH. You can use either benchtop UV lamps or light sources installed onmicroscopes. 

The calibration of the photoactivation
The objective is to provide enough photons to exhaust the conversion of the Actiflash but without generating detrimental side-effects on the biological sample.Simply analyze the phenotype recovery with decreasing illumination duration. Then determine the shortest illumination duration leading to 100% uncaging of Actiflash.

Actiflash for Zebrafish: it is efficient with zebrafish embryos of less than 48 hpf. It does not work over 48 hpf.

Storage 

Technical information

One 5mg vial
The volume of DMSO required to reach a concentration of 10mM is indicated on the vial

Want more?
We can also prepare a 50mg vial.  Ask us  for this special mice quote. 

Actiflash FAQ

Actiflash  Safety Datasheet

Actiflash  Certificate of Analysis

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Actiflash & plasmids

These 52 plasmids were developed by Sophie Vriz and Michel Volovitch in The Sophie Vriz Lab, who have the ownership of citations related to these plasmids.
These 52 plasmids are all available on Addgene.

Type Addgene Vriz lab name & link to Addgene Coding sequence Main use Regulatory sequences
ERT2 ready for fusion
184057 pCSnERT(#186) nls-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
ERT2 fused to CRE
184058 pCSCRET2(#323) 6myc-CRE-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184059 pT2EFCRET(#324) CRE-ERT2 Zebrafish Expression X.laevis-EF1
184060 pT2h70CRET(#325) CRE-ERT2 Zebrafish Expression D.rerio-hsp70, SP6
184061 pT22iU66CriT(#387) CRE-ERT2 Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi, SP6
184062 pT22iD36CriT(#388) CRE-ERT2 Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Dio3
ERT2 fused to Fluorescent Protein or Fluorogen-activated peptide
184063 pCSmChnERT(#99) mCherry-nls-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184064 pCSKaedenERT(#100) Kaede-nls-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184065 pCSGFPnERT(#205) eGFP-nls-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184066 pT22UbGfpnE4PCh(#301) eGFP-nls-ERT2-P2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184067 pT2iC6Dronpa2nlsER4(#636) Dronpa2-2nls-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184068 pT2iC6FAST1nls2ER4(#638) YFAST-2nls-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
ERT2 fused to Gal4 activator
184069 pCSTG4FER3(#190) Gal4bdFF-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184070 pT22gCfpUbiG4FER4P2A(#249) Gal4bdFF-ERT2-P2A Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184071 pT2iaTubG4FER3(#940) Gal4bdFF-ERT2 Zebrafish Expression C.auratus-a1Tub
184072 pT22i2Shh24G4FER3(#1041) Gal4bdFF-ERT2 Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Shha+ABC
184073 pT26CfpSox10G4FER3(#1048) Gal4bdFF-ERT2 Zebrafish Expression, stable transgenesis (blue heart) D.rerio-Sox10
184074 pT26i4Gli8G4E3(#1254) Gal4bdFF-ERT2 Zebrafish Expression, stable transgenesis (blue heart) D.rerio-GliRE
ERT2 fused to various kinases, inhibitors, redox enzymes
184075 pCSZPKIaHAERT(#118) ZfPKAinhiba-HA-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184076 pCSZPKIamutHAERT(#119) ZfmutPKAinhiba-HA-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184077 pCSmp38ERT(#143) 5myc-ZfP38a-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184078 pT22U6CatalE4PCh(#767) CatalDeltaC-ERT2-P2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi, SP6
ERT2 fused to various vertebrate transcription factors
184079 pCSmEn2ERT(#117) 6his-myc-chkEn2-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184080 pCSmEn2SRERT(#231) 6his-myc-chkEn2(SR)-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184081 pCSmEn25EERT(#232) 6his-myc-chkEn2(5E)-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184082 pCSmEn2n4ERT(#234) 6his-myc-chkEn2(n4)-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184083 pT22gCfpUbm5En2E4PCh(#251) 5myc-chkEn2-ERT2-P2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184084 pT22UbHEn2E4PCh(#262) 3HA-chkEn2-ERT2-P2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184085 pT22UbhaEng2aE4PCh(#271) 3HA-ZfEng2a-ERT2-P2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184086 pT22UbhaEng2bE4PCh(#272) 3HA-ZfEng2b-ERT2-P2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184087 pT22Ubm5Eng2aE4PCh(#283) 5myc-ZfEng2a-ERT2-P2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184088 pT22Ubm5Eng2bE4PCh(#284) 5myc-ZfEng2b-ERT2-P2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184089 pT22Ubm5En2SRE4PCh(#302) 5myc-chkEn2(SR)-ERT2-P2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184090 pCSm5En2CER4(#457) 5myc-chkEn2(C>S)-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184091 pT22Ubm4Eng2bKRPC(#493) 4myc-ZfEng2b(WW>KK)-ERT2-P2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184092 pT22gcfUbm5En25EE4PCh(#552) 5myc-chkEn2(5E)-ERT2-P2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184093 pCSm5En2DER4(#1062) 5myc-chkEn2(C>D)-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184094 pT22cUm5En2E4P3Che(#1188) 5myc-chkEn2-ERT2-5'fP2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184095 pT22cUm5En2SE4P3Che(#1189) 5myc-chkEn2(C>S)-ERT2-5'fP2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184096 pT22i5uasm5En2E4P3Che(#1208) 5myc-chkEn2-ERT2-5'fP2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) 5xUAS, SP6
184097 pT22i5uasm5En2SE4P3Che(#1209) 5myc-chkEn2(C>S)-ERT2-5'fP2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) 5xUAS, SP6
184098 pT22i5uasH3En2SE4P3Che(#1212) 3HA-chkEn2(C>S)-ERT2-5'fP2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) 5xUAS, SP6
184099 pT22i5uasm5En25EE4P3Che(#1213) 5myc-chkEn2(5E)-ERT2-5'fP2A-mCherry Zebrafish Expression, stable transgenesis (blue eye) 5xUAS, SP6
184100 pT2iD6Hoxa13aER4(#685) ZfHoxa13a-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184101 pT2iD6Hoxa13bER4(#686) ZfHoxa13b-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
184102 pT2iU6m5Hoxa13bE4PCh(#765) 5myc-ZfHoxa13b-ERT2-P2A-mCherry Zebrafish Expression D.rerio-Ubi, SP6
184103 pCSThaP6ERT(#184) HA-MmPax6-ERT2 Mammalian Expression, mRNA in vitro synthesis sCMV, SP6
ERT2 fused to Caspase-9 (apoptosis actuators and sensors)
184104 pT2iApowriter2(#336) myr-DIAP1-5myc-tEosFP-nls'-P2A-Casp9-ERT2 Zebrafish Expression D.rerio-Ubi
184105 pT22Apowriter2(#340) myr-DIAP1-5myc-tEosFP-nls'-P2A-Casp9-ERT2 Zebrafish Expression, stable transgenesis (blue eye) D.rerio-Ubi
184106 pT2iU6Apowriter2N(#355) myr-DIAP1-5myc-tEosFP-nls'-P2A-Casp9-ERT2 Zebrafish Expression D.rerio-Ubi, SP6
184107 pT22iuasvApoWtEos3N(#486) myr-DIAP1-2nls-tEosFP-nls'-P2A-Casp9-ERT2 Zebrafish Expression, stable transgenesis (blue eye) 4xUAS
184108 pT22iuasvApoWChe(#488) myr-DIAP1-nls'-mCherry-P2A-Casp9-ERT2 Zebrafish Expression, stable transgenesis (blue eye) 4xUAS

Story

" Once upon a time, a physicist, David Bensimon, asked a chemist, Ludovic Jullien, whether he could design a caged inducer to photocontrol protein activity in living organisms. For sure! However we also needed a biologist (Sophie Vriz) to accept the challenge to validate the caged Cyclofen-OH technology. It has been a long but so nice adventure, which has involved the tight integration of the work from many talented students, postdocs, and collaborators... Thanks to all of them!"

Odoo • Text and Image Easther Graudens
New projects, Idylle
"Actiflash perfectly highlights the researchers’ need to calibrate the experimental conditions in life sciences. We wanted to initiate a global scheme on this topic and find solutions to help them get reproducible results. So, when we met Ludovic Jullien's team, it was obvious to us that Actiflash was a first step in the right direction. Their technology had already been tested and approved by a number of researchers. It was then a question of making it a stable product, of producing it and disseminating it to the research communities. It is a great satisfaction to propose it today and we are very proud that Ludovic Jullien's team has chosen to widespread its technology with Idylle! Ludovic has extensive experience in transfer options, and his choice also reinforces our approach of promoting the know-how of researchers!"

Combining Actiflash

You may also like to combine Actiflash with other R&D products

Stampwell V-shape

Combine Actiflash and the Stampwell V-shape to immobilize and parallelize organoids for cell tracking

Stampwell Larvae 2

Combine Actiflash and the Stampwell Larvae 2 to immobilize and parallelize embryos for in vivo cell tracking

Contact us if you want to discuss your illumination project . Or just share insights on a research project.