Electroosmotic Flow-Based Nanoinjection Technique Using a Nanopipette for Green Microalgae.

Area of Science:

• Biotechnology
• Synthetic Biology
• Microalgal Research

Background:

• Microalgae are valuable for CO₂ fixation and biomaterial production.
• Efficient gene modification technologies are crucial for microalgal metabolic engineering.
• A versatile material delivery method is needed for diverse microalgal species.

Purpose of the Study:

• To develop a novel, efficient, and broadly applicable nanoinjection technique for single microalgal cells.
• To establish optimal conditions for material delivery into microalgae.
• To enable advanced genome editing and metabolic engineering in microalgae.

Main Methods:

• Developed a nanoinjection technique using electroosmotic flow through a nanopipette.
• Automated delivery of femtoliter-scale volumes into single cells.
• Optimized injection conditions (cell morphology, voltage, time) using FITC-dextran in Haematococcus sp. and Tetraselmis sp.

Main Results:

• Achieved injection efficiencies of 44% in Haematococcus sp. and 45% in Tetraselmis sp.
• Demonstrated precise and automated delivery at the femtoliter scale.
• Identified optimal parameters for successful nanoinjection across different microalgal species.

Conclusions:

• The novel nanoinjection technique offers a powerful tool for microalgal research.
• This method facilitates efficient genome editing and metabolic engineering.
• The technique has substantial potential for diverse microalgae applications in biomaterial production and CO₂ fixation.