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Gaussia luciferase

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Gaussia luciferase is a naturally occurring bioluminescent protein derived from the marine copepod Gaussia princeps. It catalyzes the oxidation of the substrate coelenterazine, resulting in the emission of light. This enzyme is commonly used as a reporter gene for monitoring gene expression and various biological processes in living cells and organisms.

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Lab products found in correlation

Secrete pair dual luminescence assay kit, by GeneCopoeia (3 mentions) Quickchange 2 xl site directed mutagenesis kit, by Agilent Technologies (2 mentions) Sanger sequencing, by Genewiz (2 mentions) Chemidoc imaging system, by Bio-Rad (2 mentions) Biolux gaussia luciferase assay kit, by New England Biolabs (2 mentions) Ab23750, by Abcam (2 mentions) Accel sirnas, by Horizon Discovery (2 mentions) Hpa002926, by Merck Group (2 mentions) Plus reagent, by Thermo Fisher Scientific (1 mentions) Lipofectamine ltx reagent, by Thermo Fisher Scientific (1 mentions) Lipofectamine ltx reagent with plus reagent, by Thermo Fisher Scientific (1 mentions) Anti ha tag antibody, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

Secrete-Pair Gaussia Luciferase Assay Kit (35 citations) Gaussia Luciferase Assay Kit (4 citations) Gaussia luciferase (GLuc) (3 citations)

5 protocols using gaussia luciferase

1

Mutating FOXO1 Binding Site in ALPL Promoter

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The pEZX-LvPG04 backbone plasmid contains the 1353 base pairs upstream of the ATG start site of the human ALPL gene (NM_000478) and this promoter sequence drives expression of Gaussia luciferase (Genecopoeia). QuickChange II XL Site-Directed Mutagenesis Kit (Agilent) was used to substitute the putative FOXO1 recognition site TGTTG with TATTA using forward primer: 5’-TCTGTCTCTGTGTCTGTTAATATATCTGG CTTTCTCTGGGTC-3’ and reverse primer: 5’-GACCCAGAGAAAGCCAGATATATTAACAG ACACAGAGACAGA-3’ according to manufacturer’s protocol. Mutant plasmids were selected and confirmed by Sanger sequencing (Genewiz) and transfected into cells using the Lipofectamine LTX Reagent with PLUS Reagent (Invitrogen) according to manufacturer’s protocols. Luciferase was quantified using the Secrete-Pair Dual Luminescence Assay Kit (Genecopoeia) according to manufacturer’s protocols and normalized to SEAP secretion.
Moorhead WJ I.I.I., Chu C.C., Cuevas R.A., Callahan J I.V., Wong R., Regan C., Boufford C.K., Sur S., Liu M., Gomez D., MacTaggart J.N., Kamenskiy A., Boehm M, & St. Hilaire C. (2020). Dysregulation of FOXO1 (Forkhead Box O1 Protein) Drives Calcification in Arterial Calcification due to Deficiency of CD73 and Is Present in Peripheral Artery Disease. Arteriosclerosis, Thrombosis, and Vascular Biology, 40(7), 1680-1694.
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2

Evaluating KLF4 Regulation of Gaussia Luciferase

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A KLF4 promoter-reporter plasmid that expresses the secretable Gaussia Luciferase was purchased from Genecopoeia, and transfected into plated MOVAS cells. 24 hours post-transfection, siRNAs targeting either KLF4 or non-target control were added to MOVAS cells according to manufacturer’s protocol (Dharmacon Accel siRNAs). 72 hours post-siRNA treatment, media was replaced with serum-free media containing recombinant proteins or tumor conditioned media. 16 hours after protein-containing treatment, media was removed and assayed for luciferase activity using the BioLux Gaussia Luciferase Assay Kit (New England BioLabs). Reporter plasmid activity is reported in luminescence as compared to empty plasmid control. Western Blots were also performed after TCM treatment of KLF4 knocked down vSMCs to probe for KLF4 (HPA002926, Sigma) and fibronectin (ab23750, Abcam), and were imaged using the BioRad ChemiDoc imaging system.
Murgai M., Ju W., Eason M., Kline J., Beury D., Kaczanowska S., Miettinen M.M., Kruhlak M., Lei H., Shern J.F., Cherepanova O.A., Owens G.K, & Kaplan R.N. (2017). KLF4-dependent perivascular cell plasticity mediates pre-metastatic niche formation and metastasis. Nature medicine, 23(10), 1176-1190.
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3

Evaluating KLF4 Regulation of Gaussia Luciferase

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A KLF4 promoter-reporter plasmid that expresses the secretable Gaussia Luciferase was purchased from Genecopoeia, and transfected into plated MOVAS cells. 24 hours post-transfection, siRNAs targeting either KLF4 or non-target control were added to MOVAS cells according to manufacturer’s protocol (Dharmacon Accel siRNAs). 72 hours post-siRNA treatment, media was replaced with serum-free media containing recombinant proteins or tumor conditioned media. 16 hours after protein-containing treatment, media was removed and assayed for luciferase activity using the BioLux Gaussia Luciferase Assay Kit (New England BioLabs). Reporter plasmid activity is reported in luminescence as compared to empty plasmid control. Western Blots were also performed after TCM treatment of KLF4 knocked down vSMCs to probe for KLF4 (HPA002926, Sigma) and fibronectin (ab23750, Abcam), and were imaged using the BioRad ChemiDoc imaging system.
Murgai M., Ju W., Eason M., Kline J., Beury D., Kaczanowska S., Miettinen M.M., Kruhlak M., Lei H., Shern J.F., Cherepanova O.A., Owens G.K, & Kaplan R.N. (2017). KLF4-dependent perivascular cell plasticity mediates pre-metastatic niche formation and metastasis. Nature medicine, 23(10), 1176-1190.
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4

ALPL Promoter-Driven Gaussia Luciferase

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The pEZX-LvPG04 backbone plasmid contains the 1353 base pairs upstream of the ATG start site of the human ALPL gene (NM_000478) and this promoter sequence drives expression of Gaussia luciferase (Genecopoeia). QuickChange II XL Site-Directed Mutagenesis Kit (Agilent) was used to substitute the putative FOXO1 (forkhead box O1 protein) recognition site TGTTG with TATTA using forward primer: 5’-TCTGTCTCTGTGTCTGTTAATATATCTGG CTTTCTCTGGGTC-3’ and reverse primer: 5’-GACCCAGAGAAAGCCAGATATATTAACAG ACACAGAGACAGA-3’ according to manufacturer’s protocol. Mutant plasmids were selected and confirmed by Sanger sequencing (Genewiz) and transfected into cells using the Lipofectamine LTX Reagent with PLUS Reagent (Invitrogen) according to manufacturer’s protocols. Luciferase was quantified using the Secrete-Pair Dual Luminescence Assay Kit (Genecopoeia) according to manufacturer’s protocols and normalized to SEAP secretion.
Moorhead WJ I.I.I., Chu C.C., Cuevas R.A., Callahan J I.V., Wong R., Regan C., Boufford C.K., Sur S., Liu M., Gomez D., MacTaggart J.N., Kamenskiy A., Boehm M, & St. Hilaire C. (2020). Dysregulation of FOXO1 (Forkhead Box O1 Protein) Drives Calcification in Arterial Calcification due to Deficiency of CD73 and Is Present in Peripheral Artery Disease. Arteriosclerosis, Thrombosis, and Vascular Biology, 40(7), 1680-1694.
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5

Luciferase Reporter Assay of NTRK3 and ASCL1 Promoter Activity

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Exponentially growing HEK293T cells were co-transfected with either (HA)-EWSR1-WT1 (−KTS), (HA)-EWSR1-FLI1 or pcDNA3.1 expression constructs, and Gaussia Luciferase-fused NTRK3 and ASCL1 promoter pEZX-PG04 plasmids (Genecopeia). The reporter assay was performed using the Secrete-Pair Dual Luminescence Assay Kit (Genecopeia) according to the manufacturer’s instructions. Briefly, 48 hours post-transfection, growth media was collected and transferred to a luminescence-compatible microtiter plate. Secreted luciferase activity was assayed on a BioTek luminometer with XYs integration. Secreted alkaline phosphatase was use for normalization. Luciferase and alkaline phosphatase measurements were performed in triplicates and all experiments were repeated at least twice. Expression levels of HA-EWSR1-WT1 and (HA)-EWSR1-FLI1 were assayed via western blot with a rabbit monoclonal anti-HA-tag antibody (Cell Signaling Technology).
Ogura K., Somwar R., Hmeljak J., Magnan H., Benayed R., Boroujeni A.M., Bowman A.S., Mattar M.S., Khodos I., de Stanchina E., Jungbluth A., Asher M., Odintsov I., Hartono A.B., LaQuaglia M.P., Slotkin E., Pratilas C.A., Lee S.B., Spraggon L, & Ladanyi M. (2020). Therapeutic potential of NTRK3 inhibition in desmoplastic small round cell tumor. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(4), 1184-1194.
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