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Table 4 Delivery methods for transferring synthetic regulatory RNAs to bacteria

From: Natural antisense RNAs as mRNA regulatory elements in bacteria: a review on function and applications

Delivery methods Traits
Biological methods
 Transduction A specific DNA donor is required for DNA transfer to recipient bacteria
 Conjugation Requiring physical contact of recipient and donor (host strain) with a conjugative plasmid or participation of a third bacterium with a helper plasmid
Not useful for large-scale delivery applications
 Gene transformation Limited to a few naturally competent groups
Physical methods
 Electroporation Highly efficient but requires low ionic strength medium and high voltage
Not useful for large-scale delivery applications
 Laser irradiation Employs a laser to change cell permeability to allow transferal; requires physical contact of laser and cells
 Ultrasound DNA delivery (UDD); Sonoporation Appropriate approach for plasmid or DNA fragment transferal to eukaryotic cells (e.g., gene therapy)
 Heat shock transfer Mostly used for E. coli (in parallel with the calcium phosphate method)
Chemical methods
 Protein & Peptides Introduced in the late 1950s, this technique originally used high salt concentration and polycationic proteins to enhance nucleic acid entry into the cell.
Now cationic peptides are using to enhance nucleic acid delivery. Cationic peptides have been found useful for enhancing cellular uptake and/or cell targeting oligonucleotide analogs. These peptides are synthetically conjugated, used as non-covalent complexes, or used in combination with polymer formulation techniques
 Calcium phosphate Simple, effective and still widely used for nucleic acid delivery
 Artificial lipids DNA has been successfully complexed with cationic, anionic and neutral liposomes. These complexes can be handled easily, but lipid-based systems generally have significant drawbacks, including the lack of targeting and variations arising during fabrication
 Naonparticles Using carbon nanotubes, nucleic acid is delivered into cells. Magneto-transformation has also been used for nucleic acid transfer, but in that method, pulsed magnetic fields assisted the delivery of DNA using magnetic nanoparticles.