Luciferase Plasmid Guide
Luciferase is the enzyme responsible for the bioluminescence found in a diverse number of organisms, ranging from bacteria to aquatic animals to insects. Luciferase catalyzes the oxidation of a substrate, leading to the emission of photons. The robust signal, sensitivity, wide dynamic range, and resistance to photobleaching has made luciferase a common choice in assays ranging from use as a reporter gene in vitro and in vivo to use in biosensors.
The most common types of luciferase found in Addgene’s collection are Firefly luciferase (Fluc), Renilla luciferase (Rluc), and NanoLuc® Luciferase (Nluc). Luc2 is the second-generation version of Firefly luciferase that has been codon optimized for expression in mammalian systems. Other luciferases in our collection include Gaussia luciferase (Gluc) and the red and green Click-beetle luciferases (CBRluc and CBGluc, respectively). Like Firefly luciferase, Click-beetle red luciferase has also been codon optimized (CBR2opt) for mammalian expression systems.
Each type of luciferase has advantages and disadvantages depending on the application. Firefly luciferase is considerably brighter than Renilla luciferase, but is more susceptible to enzyme inhibition. Both Firefly luciferase and Renilla luciferase have short protein half-lives, making them useful as transcriptional reporters. However, they are not secreted, so cells must be lysed or the substrate must be delivered intracellularly in order to measure luminescence. Gaussia luciferase is secreted and more stable, making time course experiments possible, but its short luminescence half-life makes it too dim for many experimental systems. Firefly and Renilla luciferase are less-than-ideal protein tags due to their large size. NanoLuc® Luciferase is small, bright, and a secreted version of the protein is available - these attributes have made it popular with scientists since its release.
Luciferase-containing plasmids are commonly used to investigate the effect of regulatory elements, such as promoters, enhancers and untranslated regions, or the effect of mutations of these regulatory elements on gene expression. Empty backbones for inserting regulatory elements before or after the luciferase ORF can be found in a table below, as well as plasmids that already contain a region of interest before or after luciferase. In addition, Addgene has many luciferase expression constructs available that are driven by a strong constitutive promoter and can be used to monitor transfection or viral infection efficiency, to study changes in cell physiology, or as a reference level of expression. Scientists have developed a wide range of luciferase-based plasmid systems for many different applications. Browse the Collection Highlights section to learn more.
Highlights of Luciferase Tools in our Collection
Modified/Novel Luciferases
- AkaLuc: A modified firefly luciferase engineered to use the red-shifted substrate Akalumine with significantly brighter signal and emission in the near-infrared range, enabling enhanced bioluminescence imaging in vivo.
- Luciferases with tunable wavelengths: NanoLuc® with either SNAP‐tag or HaloTag7 tag in which labeling of the self‐labeling tag with a fluorophore shifts the emission maximum of NanoLuc to that of the fluorophore.
- Conformationally destabilized Firefly luciferase mutants: A set of three sensors that are increasingly structurally destabilized versions of Firefly luciferase which serves as a convenient toolkit to assess the proteostasis status in a wide range of experimental systems.
- LumiLuc and LumiScarlet: LumiLuc and LumiScarlet are ATP-independent luciferases derived from teLuc that emit yellow/green light and red light, respectively, for increased usefulness in in vivo applications.
Split-Luciferase and Protein-Protein Interaction Assays
- Cluc-FKBP, FRB-Nluc: Split Firefly luciferase to assess FKBP and FRP interactions in the mTOR signaling pathway in vivo.
- DULIP (DUal Luminescence-based Co-Immunoprecipitation) plasmids: Luminescence-based protein-protein screening method for the systematic and quantitative analysis of co-immunoprecipitations using both Renilla and Firefly luciferase. Plasmids are Gateway cloning compatible. Learn more about DULIP in our 27 Hot Plasmids from 2016 blog post.
- LuTHy plasmids (Double‐Readout Luminescence‐based Two‐Hybrid): Combines in vivo BRET with ex vivo luminescence-based co-immunoprecipitation to assess protein-protein interactions.
- NanoLuc® complementation: Protein-protein assays based on a split NanoLuc® have been used to assess GPCR-G protein interactions, protein dimerization, endosomal escape of cell-penetrating peptides, and more.
- nLuc/cLuc for split Firefly Luciferase complementation in plants.
Bioluminescence Resonance Energy Transfer (BRET)
- CalFlux VNT: An intracellular calcium sensor created by fusing NanoLuc® and Venus fluorophore to Troponin C Ca++ binding domain.
- Nano-lanterns, Enhanced Nano-lanterns, and Antares: Fluorescent proteins are fused to Renilla luciferase (Nano-lanterns) or NanoLuc® (Antares and Enhanced Nano-lanterns) to allow for BRET-based in vivo imaging of multiple cellular processes, biosensors, or optogenetic tools.
- pcDNA3.1+-pHlash, pRSETb-pHlash: A pH sensor created by fusing a bright mutant of Renilla luciferase (Rluc8) through an Ala-Glu-Leu linker to the circularly permuted Venus fluorophore. Learn more about pHlash sensor in our 28 Hot Plasmid Technologies from 2015 blog post.
RNA Biology
- CMV-LUC2CP/intron/ARE, CMV-LUC2CP/ARE: A splicing reporter that contains an intron in the middle of the Firefly luciferase gene. Learn more about CMV-LUC2CP/intron/ARE and CMV-LUC2CP/ARE in our 28 Hot Plasmids from 2015 blog post.
- TREAT (3(Three)′-RNA End Accumulation during Turnover): Single-molecule mRNA turnover biosensor that utilizes Renilla luciferase and a Halo tag to assay mRNA decay in real time.
- pKC-4.04, pKC-4.06: Nonsense-mediated decay reporter and control plasmid utilizing Firefly luciferase.
Transcriptional Element Screening
- Optimized STARR-seq (Self-Transcribing Active Regulatory Region) plasmids: Genome-wide screening of enhancer activity using a Firefly luciferase reporter. These plasmids improve over the original STARR-seq plasmids by using the bacterial ORI as the core promoter, which was a source of false positives with the original STARR-seq method. Learn more about STARR-seq in our 27 Hot Plasmids from 2016 blog post.
- pGL3-lRF, pGL3-LRR: Genomic tiles spanning a region of interest are cloned into a Firefly luciferase reporter vector via Gateway cloning to test for the presence of transcriptional regulatory region in specific cell types. Used to identify functional SNPs from loci determined to be important in disease.
Empty Backbones
This table is a collection of empty backbone plasmids into which you can clone your regulatory element or gene of interest into to create a luciferase reporter. Unless otherwise noted, these plasmids are meant for expression in mammalian systems.
| ID | Plasmid | Luciferase Type(s) | Description | PI | |
|---|---|---|---|---|---|
| 16539 | pBV-Luc | Firefly | Insertion of 5' promoter/enhancer regions | Bert Vogelstein | |
| 12178 | pIS0 | Firefly | Insertion of target sequence in the 3' UTR | David Bartel | |
| 12179 | pIS1 | Renilla | Insertion of target sequence in the 3' UTR | David Bartel | |
| 12177 | pIS2 | Renilla | Insertion of target sequence in the 3' UTR | David Bartel | |
| 101139 | pCMV-IRES-Renilla Luciferase-IRES-Gateway-Firefly Luciferase (pIRIGF) | Firefly/Renilla | Creation ORF-Firefly luciferase gene fusions. Renilla luciferase under the control of a CMV promoter is present for normalization | William Kaelin | |
| 71248 | pXPG | Firefly | Insertion of 5' promoter/enhancer regions. Mutations within the luciferase gene allow for increased copy number in E. coli and expression of luciferase in the cytoplasm of mammalian cells | Peter Cockerill | |
| 64784 | pGL3-Basic-IRES | Firefly | Insertion of 5' promoter/enhancer regions | Joshua Mendell | |
| 60323 | pGL4.23-GW | Firefly | Insertion of 5' promoter/enhancer regions using Gateway cloning. Lentiviral. | Jorge Ferrer | |
| 87067 | pcDNA3.1-ccdB-Nanoluc | NanoLuc® | Creation of N-terminal Nanoluc fusions using Gateway cloning | Mikko Taipale | |
| 87075 | pLenti6.2-ccdB-Nanoluc | NanoLuc® | Creation of C-terminal Nanoluc fusions using Gateway cloning. Lentival. | Mikko Taipale | |
| 87078 | pLenti6.2-Nanoluc-ccdB | NanoLuc® | Creation of N-terminal Nanoluc fusions using Gateway cloning. Lentiviral. | Mikko Taipale | |
| 12563 | PCD FL0X | Firefly | Insertion of imperfect miRNA sequence in the 3' UTR | Carl Novina | |
| 33154 | pAC-Luc | Firefly | Creation of N-terminal Firefly fusions in Drosopholia | Michael Rosbash | |
| 128046 | pGL3basic luciferase and renilla_polyA | Firefly/Renilla | Insertion of 5' promoter/enhancer regions upstream of Firefly luciferase. Renilla luciferase is co-expressed. | Oskar Laur | |
| 87070 | pcDNA3.1-Nanoluc-ccdB | NanoLuc® | Creation of N-terminal Nanoluc fusions using Gateway cloning | Mikko Taipale | |
| 87065 | pcDNA3.1-ccdB-Renilla | Renilla | Creation of C-terminal Renilla luciferase fusions using Gateway cloning | Mikko Taipale | |
| 87066 | pcDNA3.1-ccdB-Firefly | Firefly | Creation of C-terminal Firefly luciferase fusions using Gateway cloning | Mikko Taipale | |
| 87068 | pcDNA3.1-Renilla-ccdB | Renilla | Creation of N-terminal Renilla luciferase fusions using Gateway cloning | Mikko Taipale | |
| 87069 | pcDNA3.1-Firefly-ccdB | Firefly | Creation of N-terminal Firefly luciferase fusions using Gateway cloning | Mikko Taipale | |
| 87074 | pLenti6.2-ccdB-Firefly | Firefly | Creation of C-terminal Firefly luciferase fusions using Gateway cloning. Lentiviral. | Mikko Taipale | |
| 87077 | pLenti6.2-Firefly-ccdB | Firefly | Creation of N-terminal Firefly luciferase fusions using Gateway cloning. Lentiviral. | Mikko Taipale | |
| 87789 | pPTluc | Firefly | Insertion of 5' promoter/enchancer regions in Drosopholia | Herman Wijnen | |
| 126033 | pTETRIS-cargo | Firefly | For use as a reporter of the ability of a given lncRNA to cis-repress | Mauro Calabrese | |
| 106253 | pLS-mP-Luc | Firefly | Insertion of 5' promoter/enhancer regions. Lentiviral | Nadav Ahituv | |
| 106292 | pLS-SV40-mP-Rluc | Renilla | Insertion of 5' promoter/enhancer regions. Lentiviral | Nadav Ahituv | |
| 108228 | pGreen2x35SLUC | Firefly/Renilla | Creation ORF-Firefly luciferase gene fusions in plants. Renilla luciferase under the control of a CMV promoter is present for normalization | Alejandro Ferrando | |
| 114670 | pGL3-TK-5UTR-BsmBI-Luciferase | Firefly | Insertion of 5' promoter/enhancer regions. Blue/white screening can be used for screening of transformants in cloning process. | Lynne Postovit | |
| 119760 | pSGDlucV3.0 | Firefly/Renilla | Insertion of 5' promoter/enhancer regions upstream of Firefly luciferase. Renilla luciferase is expressed from a T7 promoter for normalization. FMDV StopGo sequences flank the MCS for improved normalization. | John Atkins | |
| 167576 | mpCHECK2 | Firefly/Renilla | Insertion of 3' UTR sequences downstream of Renilla luciferase. Firefly luciferase expression under the control of a HSV TK promoter for normalization | Ting Ni | |
| 174050 | pGWB-nLUC | Firefly | Creation of N-terminal Firefly luciferase fragment for split-luciferase complementation assay in plants using Gateway cloning. Use with 174051 | Alberto Macho | |
| 174051 | pGWB-cLUC | Firefly | Creation of C-terminal Fifrefly luciferase fragment for split-luciferase complementation assay in plants using Gateway cloning. Use with 174050 | Alberto Macho | |
| 136010 | psi-CHECK3 | Firefly/Renilla | Insertion of 3' UTR sequences downstream of Renilla luciferase. Firefly luciferase expression under the control of a HSV TK promoter for normalization. Modified from psi-CHECK2 so both Renilla and Firefly luciferase contain introns | Anthony Leung | |
| 141285 | pGWB401NL3F10H | NanoLuc® | Creation of C-terminal luciferase fusions (NanoLuc-3xFLAG-10xHis) using Gateway cloning for expression in plants | Andrew Millar | |
| 141285 | pGWB501NL3F10H | NanoLuc® | Creation of C-terminal luciferase fusions (NanoLuc-3xFLAG-10xHis) using Gateway cloning for expression in plants | Andrew Millar | |
| 141285 | pGWB601NL3F10H | NanoLuc® | Creation of C-terminal luciferase fusions (NanoLuc-3xFLAG-10xHis) using Gateway cloning for expression in plants | Andrew Millar | |
| 141285 | pGWB701NL3F10H | NanoLuc® | Creation of C-terminal luciferase fusions (NanoLuc-3xFLAG-10xHis) using Gateway cloning for expression in plants | Andrew Millar | |
| 159707 | pG20_LUC2_Hyg | Firefly | Creation of C-terminal Firefly luciferase fusion for expression in plants | Hans-Henning Kunz | |
| 159708 | pG20_LUC2_Blp | Firefly | Creation of C-terminal Firefly luciferase fusion for expression in plants | Hans-Henning Kunz | |
| 141305 | pRATIO2251 | Firefly | Creation of C-terminal Firefly luciferase fusion for expression in plants, co-expressed with mScarlet-I for ratiometric imaging | David Nelson | |
| 141301 | pRATIO1267 | RedLuc/Gaussia | Creation of C-terminal Red Luciferase fusion for expression in plants, co-expressed with Gaussia luciferase for ratiometric imaging | David Nelson |
Expression Constructs
Luciferase driven by a strong constitutive promoter and can be used to monitor transfection or viral infection efficiency, to study changes in cell physiology, or as a reference level of expression. Use this table to find plasmids expressing luciferase. We also offer ready-to-use AAV preparations of select luciferase expression plasmids; these are noted with the viral service icon 
.
| ID | Plasmid | Luciferase Type | Promoter | Description | PI | |
|---|---|---|---|---|---|---|
| 60226 | AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR | Renilla | EF1α | AAV9-CRISPR knockout plasmid. Renilla luciferase is used to monitor viral transfection | Feng Zhang | |
| 21471 | pLenti PGK V5-LUC Neo (w623-2) | Firefly | PGK | Lentiviral expression of firefly luciferase | Eric Campeau | |
| 21474 | pLenti CMV V5-LUC Blast (w567-1) | Firefly | CMV | Lentiviral expression of firefly luciferase | Eric Campeau | |
| 18964 | Luciferase-pcDNA3 | Firefly | CMV | Mammalian expression of firefly luciferase | William Kaelin | |
| 21375 | pHIV-Luciferase | Firefly | EF1α/IRES | Lentiviral expression of firefly luciferase and gene of interest | Bryan Welm | |
| 18760 | MSCV IRES Luciferase | Firefly | (IRES) | Retroviral expression of firefly luciferase and gene of interest | Scott Lowe | |
| 18782 | MSCV Luciferase PGK-hygro | Firefly | SV40 | Retroviral expression of firefly luciferase | Scott Lowe | |
| 14080 | pAKlux2 | Lux Operon | Bacterial expression of luciferase from broad host range plasmid | Attila Karsi | ||
| 105621 | Lenti-luciferase-P2A-Neo | Firefly | EF1α | Lentiviral expression of firefly luciferase | Christopher Vakoc | |
| 33307 | Ubc.Luc.IRES.Puro | Firefly | Ubc | Lentiviral expression of firefly luciferase | Linzhao Cheng | |
| 74444 | pLenti.PGK.blast-Renilla_Luciferase | Renilla | PGK | Lentiviral expression of renilla luciferase | Reuben Shaw | |
| 48688 | pUltra-Chili-Luc | Firefly | Ubc | Lentiviral expression of firefly luciferase and dTomato. A gene of interest can also be inserted into the plasmid. | Malcolm Moore | |
| 51819 | Syn-ATP | Firefly (modified) | CMV | Optical reporter of presynaptic ATP | Timothy Ryan | |
| 24348 | pAC-hRluc | Renilla | Actin5C | Insect expression of renilla luciferase | Liqun Luo | |
| 51970 | Nano-lantern/pcDNA3 | Nano-lantern | CMV | Mammalian expression of Nano-lantern | Takeharu Nagai | |
| 87121 | pcDNA-RLuc8 | Renilla | CMV | Mammalian expression renilla luciferase mutant Rluc8, which has increased stability and light output | Sanjiv Sam Gambhir | |
| 104587 | pHIV-iRFP720-E2A-Luc | Firefly | EF1α | Lentiviral expression of firefly luciferase and iRFP720 from a bicistronic promoter | Antonius Plagge | |
| 118017 | pLX313-Firefly luciferase | Firefly | EF1α | Lentiviral expression of firefly luciferase | William Hahn, David Root | |
| 118016 | pLX313-Renilla luciferase | Renilla | EF1α | Lentiviral expression of renilla luciferase | William Hahn, David Root | |
| 105533 | pAAV.CMV.Luc.IRES.EGFP.SV40 | Firefly | CMV | AAV expression of firefly luciferase and GFP | James Wilson | |
| 22522 | phGluc | Gaussia | EF1α | Expression of Gaussia luciferase; GFP is expressed if cells are infected with virus | Christopher Buck | |
| 98580 | pLX304 Luciferase-V5 blast | Firefly | CMV | Lentiviral expression of firefly luciferase with a V5 tag | Kevin Janes | |
| 83281 | pAAV-CAG-FLuc | Firefly | CAG | AAV expression of firefly luciferase | Mark Kay | |
| 105532 | pAAV.CMV.ffLuciferase.SV40 |
Firefly | CMV | AAV expression of firefly luciferase | James Wilson | |
| 101156 | T7-CMVtrans-FFLuc-polyA | Firefly | T7 | Expression of firefly luciferase from a cell-free mammalian cell lysate | Marcel Bruchez | |
| 108542 | pLenti-EF1a-Luciferase-IRES-Blast-WPRE | Firefly | EF1α | Lentiviral expression of firefly luciferase | Javier Alcudia | |
| 113442 | pcDNA3.1 NL | NanoLuc® | CMV | Mammalian expression of Nanoluc with a N-terminal Myc tag | Erich Wanker | |
| 124701 | pLenti-PGK-Venus-Akaluc (neo) | Akaluc | hPGK | Lentiviral expression of Venus-Akaluc | Roland Friedel | |
| 120798 | pRRL Luciferase | Firefly | hPGK | Lentiviral expression of firefly luciferase | Paul Khavari | |
| 109395 | EFS-Vector | Firefly | EF1α | AAV-CRISPR knockout plasmid. Firefly luciferase is used to moniter viral transfection | Sidi Chen | |
| 109396 | TBG-Vector | Firefly | TBG | AAV-CRISPR knockout plasmid. Firefly luciferase is used to moniter viral transfection | Sidi Chen | |
| 118412 | ssAAV-EF1a-FLuc-WPRE-HgHpA_bac_293 | Firefly | EF1α | AAV expression of firefly luciferase | Nicole Paulk | |
| 98294 | pF CAG luc IRES neo | Firefly | CAG | Lentiviral expression of firefly luciferase | Brent Stringer | |
| 102320 | FUW-RLuc-T2A-PuroR | Renilla | Ubc | Lentiviral Expression of renilla luciferase | Rudolf Jaenisch | |
| 105538 | pENN.AAV.TBG.PI.ffLuciferase.RBG | Firefly | TGB | AAV expression of firefly luciferase and GFP | James Wilson | |
| 106457 | pCR3.1-Luc | Firefly | CMV | Mammalian expression of firefly luciferase | Georg Weber | |
| 113450 | pLenti NL | NanoLuc® | Ubc | Lentiviral expression of Nanoluc with a N-terminal Myc tag | Erich Wanker | |
| 115352 | pFL-SV40 | Firefly | SV40 | Mammalian expression of firefly luciferase | Ming-Chih Lai | |
| 115366 | p CIneo-RL | Renilla | CMV | Mammalian expression of humanized renilla luciferase | Witold Filipowicz | |
| 100984 | pGL4.18 CMV-Luc | Firefly | CMV | Mammalian expression of firefly luciferase | Lee Helman | |
| 123655 | pKM19 | Firefly | pIEx | Insect expression of firefly luciferase | Kevin Maringer | |
| 131860 | pZmUBQ:LUC | Firefly | CaMV | Zea mays expression of firefly luciferase | Paul Schulze-Lefert | |
| 83282 | pAAV-CAG-RLuc | Renilla | CAG | AAV expression of renilla luciferase | Mark Kay | |
| 140328 | pLenti-PGK-Venus-Fluc (puro) | Firefly | hPGK | Lentiviral expression of Venus-firefly luciferase | Roland Friedel | |
| 170575 | pCMV-FLuc | Firefly | CMV | Retroviral expression of firefly luciferase | David Nemazee | |
| 68216 | pEGB 35S:Luciferase:Tnos (GB0110) | Firefly | 35S | Expression of firefly luciferase in plants | Diego Orzaez | |
| 68215 | pEGB 35S:Renilla:Tnos (GB0109) | Renilla | 35S | Expression of Renilla luciferase in plants | Diego Orzaez | |
| 127223 | pMOD_A5801 | Firefly | 35S | Expression of firefly luciferase in plants | Daniel Voytas | |
| 127224 | pMOD_A5803 | Firefly | CmYLCV | Expression of firefly luciferase in plants | Daniel Voytas |
Reporter Constructs
Already know what gene or regulatory element that you need a luciferase reporter for? Browse or use the search box to find a construct that contains your regulatory element of interest.
| ID | Plasmid | Description | Gene/Insert | PI |
|---|
Do you have suggestions for other plasmids that should be added to this list?
Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this resource!