Abstract
Chronic hepatitis B virus (HBV) poses a significant global health challenge as it can lead to acute or chronic liver disease and hepatocellular carcinoma (HCC). To establish a safety experimental model, a homolog of HBV—duck HBV (DHBV) is often used for HBV research. Hydrodynamic-based gene delivery (HGD) is an efficient method to introduce exogenous genes into the liver, making it suitable for basic research. In this study, a duck HGD system was first constructed by injecting the reporter plasmid pLIVE-SEAP via the ankle vein. The highest expression of SEAP occurred when ducks were injected with 5 µg/mL plasmid pLIVE-SEAP in 10% bodyweight volume of physiological saline for 6 s. To verify the distribution and expression of exogenous genes in multiple tissues, the relative level of foreign gene DNA and β-galactosidase staining of LacZ were evaluated, which showed the plasmids and their products were located mainly in the liver. Additionally, β-galactosidase staining and fluorescence imaging indicated the delivered exogenous genes could be expressed in a short time. Further, the application of the duck HGD model on DHBV treatment was investigated by transferring representative anti-HBV genes IFNα and IFNγ into DHBV-infected ducks. Delivery of plasmids expressing IFNα and IFNγ inhibited DHBV infection and we established a novel efficient HGD method in ducks, which could be useful for drug screening of new genes, mRNAs and proteins for anti-HBV treatment.
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The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- HGD:
-
Hydrodynamic gene delivery
- DHBV:
-
Duck Hepatitis B Virus
- qPCR:
-
Quantitative PCR
- SEAP:
-
Secreted embryonic alkaline phosphatase
- GFP:
-
Green fluorescent protein
- IFNα:
-
Interferon α
- IFNγ:
-
Interferon γ
- AST:
-
Aspartate transaminase
- ALT:
-
Alanine aminotransferase
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Funding
Natural Science Foundation of China (82160259, 31771400, 81860662, 81728006, 81501129), Natural Science Foundation of Jiangxi Province (20212ACB206039, 20171ACB21001, 20171BCB23029, 20161BAB215200).
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ZJZ, JBZ, LJZ, NS, XYH, and YTH—performed the experiments. ZJZ, JBZ, LJZ, KLC, and CBZ—wrote and edited the manuscript. CBZ, DRX, and HBX—supervised the study. All coauthors approved the manuscript.
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All institutional and national guidelines for the care and use of laboratory animals were followed, and they comply with the ARRIVE guidelines. The ethics committee: the Animal Ethics Committee of Nanchang University. The number or ID of the ethics approval: NCULAE-202209280017.
Considering the availability of ducks and the advantages of DHBV, we chose ducks as the experimental material. We cumulative used 60 male ducks to study the HGD system and its application. According to the “laboratory animal-Requirements of environment and housing facilities” (GB 14925-2010, Standardization Administration of the People’s Republic of China), all ducks were under the conditions of room temperature (24 ± 0.5 °C), humidity (50 ± 10%), light (12 h light/dark cycle) and were free access to diet. According to the “Technical specifications on euthanasia of laboratory animals” (RB/T 061-2021, Certification and Accreditation Administration of the People’s Republic of China), the ducks were euthanized with excessive isoflurane at the end of study.
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Zhao, Z., Zhu, J., Zhou, L. et al. Establishment of a hydrodynamic delivery system in ducks. Transgenic Res 33, 35–46 (2024). https://doi.org/10.1007/s11248-024-00377-x
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DOI: https://doi.org/10.1007/s11248-024-00377-x