Generation and application of a novel transgenic zebrafish line Tg(GAcyp1a:eGFP/Luc) as an in vivo assay to sensitive and specific monitoring of DLCs in the environment

Shaolin Xie; Bing Yang; Siying Li; Liangjun Ge; Min Li; Qingshi Chen; Xian Qing; Jixing Zou

Ecotoxicology and Environmental Safety (Oct 2023)

Generation and application of a novel transgenic zebrafish line Tg(GAcyp1a:eGFP/Luc) as an in vivo assay to sensitive and specific monitoring of DLCs in the environment

Shaolin Xie,
Bing Yang,
Siying Li,
Liangjun Ge,
Min Li,
Qingshi Chen,
Xian Qing,
Jixing Zou

Affiliations

Shaolin Xie: College of Marine Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, China; Guangdong Laboratory for Lingnan Modern Agriculture, South China Agricultural University, Guangzhou 510642, China
Bing Yang: College of Marine Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, China; CAS Key Laboratory of Tropical Marine Bio-Resources and Ecology, Guangdong Provincial Key Laboratory of Applied Marine Biology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China
Siying Li: College of Marine Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, China
Liangjun Ge: College of Marine Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, China
Min Li: College of Marine Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, China
Qingshi Chen: College of Marine Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, China
Xian Qing: South China Institute of Environmental Sciences, Ministry of Ecology and Environment, Guangzhou 510530, China; Corresponding author.
Jixing Zou: College of Marine Sciences, South China Agricultural University, Guangzhou, Guangdong 510642, China; Guangdong Laboratory for Lingnan Modern Agriculture, South China Agricultural University, Guangzhou 510642, China; Corresponding author at: College of Marine Sciences, South China Agricultural University, Guangzhou 510642, China.

Journal volume & issue: Vol. 264
p. 115471

Abstract

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CYP1A is the most commonly used biomarker and transgenic fish which carrying a cyp1a promoter to drive a reporter gene can be used as reliable way to monitor dioxin/dioxin-like compounds (DLCs) in the environment. Here, we cloned the cyp1a promoter of Gambusia affinis and this promoter showed stronger transcriptional activity than that of zebrafish. Then, a Tg(GAcyp1a:eGFP/Luc) transgenic zebrafish line was first constructed with the G. affinis cyp1a promoter driving eGFP expression using meganuclease I-SceI mediated transgenesis technology. The Tg(GAcyp1a:eGFP/Luc) larvae at 72 h post-fertilization (hpf) were tested by exposing to TCDD for 72 h, and induced GFP was mainly expressed in the liver with low background. The Tg(GAcyp1a:eGFP/Luc) zebrafish showed high sensitivity (limit of detection of 0.322 ng/L TCDD and 0.7 TEQ-ng/L PCDD/Fs) and specificity (insensitive to responses to PAHs and PCBs). In addition, the transgenic line showed a low detection concentration of the DLCs contaminated environmental samples (as low as 1.8 TEQ-ng/L), and the eGFP fluorescence intensity and the chemical-TEQ values were closely correlated. In conclusion, a sensitively and specifically transgenic zebrafish line was established to convenient and effective to detect DLCs in the environment.

Published in Ecotoxicology and Environmental Safety

ISSN: 0147-6513 (Print); 1090-2414 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Technology: Environmental technology. Sanitary engineering: Environmental pollution; Geography. Anthropology. Recreation: Environmental sciences
Website: https://www.journals.elsevier.com/ecotoxicology-and-environmental-safety

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