# A cost-effective dual reporter system in Nicotiana benthamiana

**Authors:** Zeru Zhang, Shuangle Yan, Fuqi Li, Zhijuan Rao, Fahong Fan, Haibin Ren, Yanlin Qu, Ke Yang, Chuanyu Ma, Chen Zhuang, Chentao Lin, Gaoping Qu

PMC · DOI: 10.3389/fpls.2026.1732683 · 2026-02-03

## TL;DR

The paper introduces a cost-effective dual reporter system using NanoLuc-GFP in Nicotiana benthamiana for accurate gene expression studies.

## Contribution

The study identifies NanoLuc-GFP as a superior, kit-independent dual reporter system for Nicotiana benthamiana.

## Key findings

- NanoLuc-GFP shows high stability and reduced interference from biological and environmental variability.
- The system is more sensitive than the conventional FLuc/RLuc system.
- Seven reporters were tested, with NanoLuc-GFP emerging as the optimal choice.

## Abstract

Dual reporter assay has been widely used for quantification of gene expression in various model systems. However, the assay is dependent on commercial kits, which may be inhibitory for many low-budget academic laboratories or high throughput screening. Here, we systematically characterized seven commonly used or potential reporters, and identified NanoLuc-GFP pair as the optimal kit-independent and accurate dual reporter in Nicotiana benthamiana. The NanoLuc-GFP system exhibited high stability, which can largely reduce the interference caused by biological variability and dramatic environmental fluctuations. Furthermore, it exhibits greater sensitivity compared to the conventional FLuc/RLuc system commercially available.

## Linked entities

- **Proteins:** NAL1 (Protein NARROW LEAF 1), rluC (ribosomal large subunit pseudouridine synthase C)
- **Species:** Nicotiana benthamiana (taxon 4100)

## Full-text entities

- **Chemicals:** MgCl2 (MESH:D015636), DMF (-), gold (MESH:D006046), NaCl (MESH:D012965), CTZ (MESH:C017144), glycerol (MESH:D005990), Furimazine (MESH:C000713648), EDTA (MESH:D004492), Triton X-100 (MESH:D017830), coenzyme A (MESH:D003065), chlorophyll (MESH:D002734), acetosyringone (MESH:C051667), ATP (MESH:D000255), coelenteramide (MESH:C542661), luciferin (MESH:D000090562), dehydroluciferyl adenylate (MESH:C107793), MES (MESH:C004550), phenylbenzothiazole (MESH:C070845), ethanol (MESH:D000431), oxyluciferin (MESH:C014730), DMSO (MESH:D004121)
- **Species:** Escherichia coli (E. coli, species) [taxon 562], Bean yellow dwarf virus (no rank) [taxon 57119], Saccharomyces cerevisiae (baker's yeast, species) [taxon 4932], Oplophorus gracilirostris (species) [taxon 727944], Nicotiana benthamiana (species) [taxon 4100], Renilla reniformis (sea pansy, species) [taxon 6136], Nicotiana tabacum (American tobacco, species) [taxon 4097], Agrobacterium tumefaciens (species) [taxon 358], Photinus pyralis (common eastern firefly, species) [taxon 7054]
- **Cell lines:** HEK293T — Homo sapiens (Human), Transformed cell line (CVCL_0063), C58C1 — Oncorhynchus mykiss (Rainbow trout), Spontaneously immortalized cell line (CVCL_S157)

## Figures

6 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12909557/full.md

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Source: https://tomesphere.com/paper/PMC12909557