Efficient gene delivery into fish cells by an improved recombinant baculovirus

Area of Science:

• * Molecular Biology
• * Gene Therapy
• * Aquaculture

Background:

• * Baculoviruses are effective gene delivery vectors in mammalian cells.
• * Previous attempts showed low gene delivery efficiency in differentiated fish cells.

Purpose of the Study:

• * To construct an improved recombinant baculovirus for enhanced gene delivery in fish cells.
• * To evaluate the transduction efficiency and transgene persistence in fish cell lines.

Main Methods:

• * Construction of a recombinant baculovirus with cytomegalovirus immediate-early (CMV-IE) promoter and enhanced green fluorescent protein (EGFP).
• * Transduction of fish cell lines using the improved baculovirus vector.
• * Measurement of transduction efficiency via flow cytometry (FCM) and monitoring EGFP persistence using fluorescence microscopy.

Main Results:

• * High-efficiency gene delivery into differentiated fish cells was achieved using the improved baculovirus vector.
• * Growth medium outperformed PBS in enhancing transduction efficiency for certain fish cells.
• * Transgene expression (EGFP) showed prolonged persistence in fish cells, peaking several days post-transduction.

Conclusions:

• * The improved recombinant baculovirus is an excellent gene delivery vector for fish cells.
• * This study provides new insights into baculovirus transduction of vertebrate cells.
• * The findings support the potential application of this vector in fish cell research and biotechnology.