Higher intensity luciferase
Akaluc luciferase providing brighter and red-shifted luminescence
Firefly bioluminescence systems have been commonly used as a imaging tool of biological phenomena. However, it have not been strong enough for imaging signals in deep tissues, because of low permeabilization of substrates. Recently, Dr. Atsushi Miyawaki and Dr. Satoshi Iwano of the RIKEN Center for Brain Science, and Dr. Shojiro Maki of the University of Electro-Communications developed a system of artificial bioluminescence AkaBLI that enables noninvasive signal observation in deep tissue of living animals.
The AkaBLI system consists of an artificial substrate AkaLumine with improved tissue permeability and an artificial luciferase Akaluc optimized to AkaLumine. The intensity of the luminescence of AkaBLI system is 100 to 1000 folds brighter than the conventional systems.
The expression vector of artificial luciferase Akaluc has been deposited by Dr. Miyawaki’s lab and it is now available from the DNA Bank.
Iwano, S. et al., Single-cell bioluminescence imaging of deep tissue in freely moving animals. Science 359 (6378): 935-939 (2018). PMID 29472486.
- Press Release and Blog
In living color: seeing cells from outside the body with synthetic bioluminescence (2018/2/23, RIKEN)
In living color: imaging the brain with synthetic bioluminescence (2018/2/23, It Ain’t Magic Blog)
- DNA resource
pcDNA3 Venus-Akaluc (cat# RDB15781)
pAAV2 SynTetOff Venus-Akaluc (cat# RDB15782)
pAAV2 TRE Venus-Akaluc (cat# RDB15783)
Brighter than commonly used luciferase!
These luciferases have at most four-time maximum luminous intensity than that of widely used luciferase of the Photinus pyralis, a common North American firefly. These luciferases as a reporter, allow to monitor gene expression with high sensitivity and time-lapse observation.
Observation of luminescence of transfected HeLa cells.
HeLa cells were transfected with expression vectors harbouring luciferase coding regions, respectively. Luminescence was observed under the condition of 2 mM D-luciferin. Left, PmatLuc1, DkumLuc1, MALuci2Luc. Right, SflaRE1Luc, PsagRE1Luc.
|Catalog no.||Name of clone||Origin||Intracellular luminescence intensity(vie, Phothinus pyralis)||Maximal absorption wavelength (pH8, in vitro)||misc.|
|RDB14359||pPmatLuc1||Pyrocoeli matsumurai||4 times or more||560 nm||567 nm||luciferase cDNA is cloned in pUC19 vector.|
|RDB14360||pDkumLuc1||Drilaster kumejimensis||4 times||558 nm||562 nm|
|RDB14363||pMALuci2Luc||Malaysian Luciola sp. (1)||2 times||556 nm||557 nm|
|RDB14362||pSflaRE1Luc||Stenocladius flavipennis||2 times||608 nm||609 nm|
|RDB14361||pPsagRE1Luc||Pyrocoeli sagulatus||2 times||605 nm||606 nm|
1The luciferase gene originated from Malaysian firefly was developed together with Perak State Development Corporation and Nimura Genetic Solutions in conformity with the Convention on Biological Diversity.
Multiple colors with one substrate!
Green-, Yellow- or Red-emission can be obtained by using D-luciferin as a substrate of each luciferase. Simultaneous monitoring of two genes can be allowed by using green- and red-luciferases.
High stabilities under various pH conditions!
These luciferases are less susceptible under variety of pH or temperature and the stable results can be obtained. Please visit our web site to see results of luminescence at 25℃ and 37℃.
|Purified recombinant luciferases were mixed with GTA Buffer (final conc. 200 mM), ATP (final conc. 2 mM), MgSO4 (final conc. 2 mM), D-Luciferin (final conc. 1 mM) and luminescence under the different pH conditions were observed. Left, 25℃. Right, 37℃.|
Akiyoshi R, Ogoh K, Suzuki H. 2012. Firefly luciferase. Japan patent unregistered 2012-235756
Akiyoshi R, Ogoh K, Suzuki H. 2013. Firefly luciferase. Japan patent unregistered 2013-74861
Ogoh K, Akiyoshi R, Suzuki H. 2013. Firefly luciferase. Japan patent unregistered 2013-138668
Ogoh K, Akiyoshi R, Suzuki H. 2013. Firefly luciferase. Japan patent unregistered 2013-81459
Akiyoshi R, Ogoh K, Suzuki H. 2015. FIREFLY LUCIFERASE, Patent publication US2015/0291938 A1
- Fluorescent protein resource
- Sea cactus therostable GFP
- Azami Green (AG, hmAG1, hmAG407)
- cjBlue (cjBlue, cjBlue Y64L)
- Dronpa-Green (Dronpa, Dronpa2, Dronpa3, 22G)
- Keima-red (dKeima, dKeima570, mKeima, tdKeima)
- Kikume Green-Red (KikGR, mKikGR, mKikGR13.2, Xpa)
- Kusabira Green Orange (mK-GO)
- Kusabira Orange (KO1, hmKO1, hmKO2, hmKO-K)
- Midoriishi-Cyan (MiCy, mMiCy1)
- Venus (Venus, mVenus, cp49Venus, cp145Venus, cp157Venus, cp173Venus, cp195Venus, cp229Venus)
- Destabilized FPs
- Knock in markers with CRISPR/Cas9 genome editing
- Nano-lantern luminescent and fluorescent protein
Sensor & Visualization
- Sensor & Visualization
- Bilirubin indicator UnaG, BReleaCa
- cAMP indicator
- Caspase activity indicator
- pH sensor
- Protein concentration in cells
- RA indicator
- Voltage indicator
- Autophagy indicator
- Chaperone mediated autophagy activity by the GAPDH-HT indicator by Dr. Takahiro Seki’s lab as well as fluorescent protein probes of LC3B accumulation developed by Dr. Noboru Mizushima’s lab, Dr. Itaru Hamachi’s lab and Dr. Keiji Kimura’s lab are available.
- Calcium-ion sensor
- Fluorescent protein-based Ca2+ sensors, G-CaMPs, Yellow Cameleons and Pericams developed by Dr. Jin-ichi Nakai’s lab and Dr. Atsushi Miyawaki’s lab, which are composed with calmodulin, fluorescent protein and M13 peptide (CaM binding domain of myosin light chain kinase), are designated to visualize intracellular [Ca2+] dynamics.
- Yellow Cameleon
- Cell cycle indicator Fucci
- To monitor cell cycle progression in living cells, cell cycle indicator Fucci probes deveoped by Dr. Atsushi Miyawaki’s lab are available.
- Epigenetics reporter
- Visualization of histone acetylation: The Histac fluorescent probes deposited by Dr. Kazuki Sasaki allow you monitoring the state of activity of acetylation of histone H4 by fluorescence in living cells.
- Visualization of methylated DNA: The EGFP-MBD-nls protein recognizes the methylated DNA and you can follow status of the DNA methylation in situ under physiological conditions using the pEGFP-MBD-nls expression clone.
- Knock in markers with CRISPR/Cas9 genome editing
- As a part of Auxin Inducible Degron (AID) System clones, Dr. Masato Kanemaki provides a series of knock-in fluorescent and selection markers with CRISPR/Cas9 genome editing.
- Notch signaling reporter
- The pRBS-EGFP and RBP-J-Venus expression clones deposited by Dr. Makoto Mark Taketo and Dr. Kenji Tanigaki, respectively, allow you monitoring the state of activation of the Notch signaling by fluorescence in living cells.
- Organelle marker/subcellular localization
- We are providing genetic resources for visualization of organelles such as mitochondria and nucleus. Each clone contains an organelle localization signal sequence fused with fluorescent proteins or epitope tags. Organelles can be detected by fluorescence or by detecting epitope tags with antibodies.
- Visualization of organelle contact sites using the organelle-targeted split-GFP system developed by Dr. Yasushi Tamura
- Sphingolipid marker
- Lipid rafts are small lipid domains on the cell membrane and are thought to play an important role in signal transduction, endocytosis and more. We provide fluorescent probes for sphingomyelin and cholesterol lipid domains.
- Nakanori: sphingomyelin and cholesterol lipid domain (lipid raft)
- D4 toxin: cholesterol rich domain
- lysenin: sphingomyelin
(GRP0025e 2016.11.16 N.N.)