Introducing foreign DNA into tiger shrimp (Penaeus monodon) by electroporation

Area of Science:

• Aquaculture
• Molecular Biology
• Genetics

Background:

• Genetic modification holds potential for improving aquaculture species like tiger shrimp (Penaeus monodon).
• Electroporation is a physical method for introducing foreign DNA into cells.

Purpose of the Study:

• To assess the feasibility of generating transgenic tiger shrimp using electroporation.
• To evaluate the efficiency and effects of DNA delivery via electroporation on shrimp development and gene expression.

Main Methods:

• Penaeus monodon zygotes were electroporated with a DNA construct (pFLAG-CMV-1-BAP) containing a bacterial alkaline phosphatase gene under a human cytomegalovirus (CMV) promoter.
• Optimization of electroporation parameters included voltage (10kV), pulse number (28), pulse time (120 μs), cycles (10), and DNA concentration (37.5 μg/mL).
• Gene transfer efficiency was assessed using dot blot and genomic Southern blotting, while protein expression was confirmed by immunohistochemical staining.

Main Results:

• Electroporation significantly reduced zygote hatching rates (46%) compared to controls (89%).
• Transgenic tiger shrimp exhibited lower postlarval survival rates (0.6% at PL45 to 0.4% at PL120).
• Gene transfer was detected in 37% of mysis-stage shrimp, with integration into the genomes of 31% of transgenic individuals, showing mosaic distribution across tissues. The FLAG-BAP protein was found in some shrimp ovaries.

Conclusions:

• Electroporation can generate transgenic tiger shrimp, but it negatively impacts early developmental stages and survival.
• The introduced DNA integrates into the shrimp genome and is expressed, though its distribution is mosaic.
• Further research is needed to optimize electroporation protocols for improved efficiency and reduced adverse effects in Penaeus monodon aquaculture.