Electrotransfection of Polyamine Folded DNA Origami Structures

Area of Science:

• Molecular nanotechnology
• Biophysics
• Synthetic biology

Background:

• DNA origami enables the creation of complex nanoscale structures.
• Conventional methods often require specific buffer conditions and may lack stability.

Purpose of the Study:

• To develop a novel method for forming DNA origami structures.
• To enhance the stability and cellular delivery of DNA origami.

Main Methods:

• Utilized spermidine as a condensing agent in magnesium-free buffer solutions.
• Investigated the formation of DNA origami sheets, tubes, and blocks.
• Assessed structural stability under high electric field pulses and in cell lysate.
• Demonstrated cellular delivery via electroporation.

Main Results:

• Successfully formed three types of DNA origami structures (sheet, tube, block) using low spermidine concentrations.
• Spermidine concentration is proportional to DNA concentration for optimal folding.
• Excessive spermidine leads to aggregation and precipitation.
• Spermidine-stabilized structures exhibit remarkable stability against high electric fields.
• Structures remain stable in cell lysate and are efficiently delivered into mammalian cells.

Conclusions:

• Spermidine offers a viable alternative to conventional buffers for DNA origami formation.
• This method yields robust DNA origami structures suitable for biological applications.
• Enhanced stability facilitates successful delivery into cells, opening avenues for nanomedicine.