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.
- Reference
Nakashiba, T. et al., Development of two mouse strains conditionally expressing bright luciferases with distinct emission spectra as new tools for in vivo imaging. Lab. Anim. (NY). 2023 Sep 7. doi: 10.1038/s41684-023-01238-6. Epub ahead of print. PMID 37679611.
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
Development of two mouse strains conditionally expressing bright luciferases with distinct emission spectra as new tools for in vivo imaging. (2023/9/8, RIKEN BRC)
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)
(2018.05.07 N.N.)
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. | |
---|---|---|---|---|---|---|
25℃ | 37℃ | |||||
RDB14359 | pPmatLuc1 | Pyrocoelia 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 | Pristolycus 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℃. | |
PmatLuc1 | |
DkumLuc1 | |
MALuci2Luc | |
PsagRE1Luc | |
SflaRE1Luc |
- Reference
Nakashiba, T. et al., Development of two mouse strains conditionally expressing bright luciferases with distinct emission spectra as new tools for in vivo imaging. Lab. Anim. (NY). 2023 Sep 7. doi: 10.1038/s41684-023-01238-6. Epub ahead of print. PMID 37679611.
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 - Press Release
Development of two mouse strains conditionally expressing bright luciferases with distinct emission spectra as new tools for in vivo imaging. (2023/9/8, RIKEN BRC)
Artificial Luciferase Variants
Catalog # | Name of clone | Running title | Publication |
---|---|---|---|
RDB19616 | ALuc23 | Expression vector of artificial luciferase variant ALuc23 derived from the copepod gene database for mammalian cell imaging. | Kim, S.B. et al., 2013 |
RDB19617 | ALuc49 | Expression vector of artificial luciferase variant ALuc49 derived from the copepod gene database for mammalian cell imaging. | Kim, S.B. et al., 2017 |
Reference
- Kim, S.B. et al. (2013) Bioconjug. Chem. 24 (12): 2067-2075. PMID 24237362
- Kim, S.B. et al. (2017) ACS Comb. Sci. 19 (9): 594-599. PMID 28742969
JNC Corporation’s Resource
Aequorea victoria Aequorin
Catalog # | Name of clone | Running title | Publication |
---|---|---|---|
RDB19836 | pAQ-B | Expression vector of A. victoria Aequorin, wild type. | Inouye, S. et al., 1985 |
RDB19830 | pJN-opAQ | Expression vector of A. victoria Aequorin, codon-optimized for human (opAQ). | Inouye, S. et al., 1985 |
RDB19847 | pJNC-opAQ | Mammalian expression vector of A. victoria Aequorin, codon-optimized (opAQ). | Inouye, S. et al., 2015 |
RDB19848 | pJNC-COsp-opAQ | Mammalian expression vector of human cytochrome C oxidase signal peptide sequence(COsp) and A. victoria Aequorin, codon-optimized (opAQ). | Inouye, S. et al., 2015 |
Clytia gregaria ClytinI and ClytinII
Catalog # | Name of clone | Running title | Publication |
---|---|---|---|
RDB19837 | pCL41 | Expression vector of C. gregaria ClytinI, wild type. | Inouye, S. et al., 1993 |
RDB19838 | pCL31 | Expression vector of C. gregaria ClytinII, wild type. | Inouye, S., 2008 |
RDB19831 | pJN-opCLII | Expression vector of C. gregaria ClytinII, codon-optimized for human (opCLII). | Inouye, S. et al., 2015 |
RDB19849 | pJNC-opCLII | Mammalian expression vector of C. gregaria ClytinII, codon-optimized (opCLII). | Inouye, S. et al., 2015 |
RDB19850 | pJNC-COsp-opCLII | Mammalian expression vector of human cytochrome C oxidase signal peptide sequence(COsp) and C. gregaria ClytinII, codon-optimized (opCLII). | Inouye, S. et al., 2015 |
Renilla reniformis luciferase
Catalog # | Name of clone | Running title | Publication |
---|---|---|---|
RDB19853 | pJNC-opRL | Mammalian expression vector of R. reniformis luciferase, codon-optimized (opRLuc). | Inouye, S. et al., 2015 |
RDB19854 | pJNC-opRL547 | Mammalian expression vector of R. reniformis Red-shiffted luciferase, codon-optimized (opRLuc547), 547 nm peak. | Unpublished bioresource |
RDB19832 | pJN-RLBP | Expression vector of R. reniformis luciferin-binding protein (RLBP). | Inouye, S. et al., 2007 |
Luciola cruciata luciferase
Catalog # | Name of clone | Running title | Publication |
---|---|---|---|
RDB19833 | pJN-opLcLuc | Expression vector of L. cruciata firefly luciferase, codon-optimized for human (opLcLuc). | Inouye, S. et al., 2015 |
Photinus pyralis luciferase
Catalog # | Name of clone | Running title | Publication |
---|---|---|---|
RDB19834 | pJN-opPyLuc | Expression vector of P. pyralis firefly luciferase, codon-optimized for human (opPyLuc). | Inouye, S. et al., 2015 |
Oplophorus gracilorostris luciferase
Catalog # | Name of clone | Running title | Publication |
---|---|---|---|
RDB19840 | pKAZ-412 | Expression vector of O. gracilorostris 19 kDa Protein of Oplophorus Luciferase (KAZ), catalytic component. | Inouye, S. et al., 2000 |
RDB19841 | pJN-eKAZ | Expression vector of O. gracilorostris enhanced 19 kDa Protein of Oplophorus Luciferase (eKAZ). | Inouye, S. et al., 2014 |
RDB19851 | pJNC-eKAZ | Mammalian expression vector of O. gracilorostris enhanced 19 kDa Protein of Oplophorus Luciferase (eKAZ). | Inouye, S. et al., 2014 |
RDB19852 | pJNC-GLsp-eKAZ | Mammalian expression vector of signal peptide sequence of G. princeps Gaussia luciferase (GLsp) and O. gracilorostris enhanced 19 kDa Protein of Oplophorus Luciferase (eKAZ). | Inouye, S. et al., 2014 |
RDB19842 | pOL23 | Expression vector of O. gracilorostris 35 kDa protein of Oplophorus luciferase, unknown function. | Inouye, S. et al., 2000 |
Gaussia princeps luciferase
Catalog # | Name of clone | Running title | Publication |
---|---|---|---|
RDB19845 | pJNC-hINS-GLuc | Mammalian expression vector of human preproinsulin (hINS) and G. princeps Gaussia luciferase (GLuc). | Suzuki, T. et al., 2011a |
RDB19846 | pJNC-hMMP2-GLuc | Mammalian expression vector of human matrix metalloprotease-2 (hMMP2) and G. princeps Gaussia luciferase (GLuc). | Suzuki, T. et al., 2011b |
RDB19855 | pJNC-pGLuc | Mammalian expression vector of G. princeps Gaussia luciferase, codon-optimized (GLuc). | Inouye, S. et al., 2016 |
Vibrio fischeri NAD(P)H-flavin oxidoreductase
Catalog # | Name of clone | Running title | Publication |
---|---|---|---|
RDB19835 | pJN-FRase | Expression vector of V. fischeri NAD(P)H-flavin oxidoreductase (FRase). | Inouye, S., 1994 |
Catalog # | Name of clone | Running title | Publication |
---|---|---|---|
RDB18951 | pHLUX20 | Expression vector of bacterial luciferase in Nitrosomonas europaea as a host strain. | Iizumi, T., 1998; Iizumi, T., 1997 |
Catalog no. | Name of resource | Type |
---|---|---|
RDB19350 | pGL4.14-E1bTATA Luciferase reporter vector of the minimal adenovirus E1b promoter sequence. |
|
RDB03988 | pCMVluc+ Expression vector of firefly luciferase |
|
RDB06411 | pGL3-RSV Expression vector of firefly luciferase, RSV promoter |
|
RDB03989 | pCAluc+ Expression vector of firefly luciferase |
|
RDB02443 | pCAcc-Luc+ Expressing luciferase under the control of CAG promoter |
|
RDB02444 | pLRICALuc+ Shuttle vector to generate recombinant adenovirus |
|
RDB02968 | pTcf7wtluc Reporter plasmid carring TCF-binding consensus sequence |
|
RDB02969 | pTcf7mtluc Reporter plasmid carring TCF-binding consensus sequence |
|
RDB07031 | ptk-luc Reporter construct with luciferase expression under tk promoter |
|
RDB07032 | pFGF4tk-luc Reporter construct with luciferase expression under FGF4 and tk promoter |
|
RDB02453 | AxCA Luc+ Recombinant adenovirus expressing luciferase gene |
|
RDB04026 | pKM2L Reporter construct with Renilla luciferase gene |
|
RDB05549 | pKM2L-pvTK HSV herpes simplex virus thymidine kinase (HSV-TK) |
|
RDB05550 | pKM2L-pvSV40 SV40 simian virus 40 early enhancer/promoter region (SV40) |
|
RDB05551 | pKM2L-pvCMV CMV cytomegalovirus immediate enhancer/promoter (CMV) |
|
RDB06408 | pRL-CMV mut1 Expression vector of Renilla luciferase, mutated CMV promoter |
|
RDB06409 | pRL-CMV mut2 Expression vector of Renilla luciferase, mutated CMV promoter |
|
RDB06410 | pRL-RSV Expression vector of Renilla luciferase, RSV promoter |
Miscellaneous
- Promoter Collection: constructs deposited by individual researchers and constructed by the Gene Engineering Division.
(GRP0025e 2016.11.16 N.N.)
2023.09.28