Nano-biolistics: a method of biolistic transfection of cells and tissues using a gene gun with novel nanometer-sized projectiles

BMC Biotechnology

Methodology Article

John A O'Brien¹  Sarah CR Lummis² 

[1] Neurobiology Division, MRC Laboratory of Molecular Biology, CB2 2QH, Cambridge, UK;Neurobiology Division, MRC Laboratory of Molecular Biology, CB2 2QH, Cambridge, UK;Department of Biochemistry, University of Cambridge, CB2 1QW, Cambridge, UK;

关键词: Gold Particle;    Spermidine;    Dendritic Spine;    Maximum Depth Penetration;    Human Embryonic Kidney HEK293;   

DOI  :  10.1186/1472-6750-11-66

 received in 2010-06-24, accepted in 2011-06-10,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundBiolistic transfection is proving an increasingly popular method of incorporating DNA or RNA into cells that are difficult to transfect using traditional methods. The technique routinely uses 'microparticles', which are ~1 μm diameter projectiles, fired into tissues using pressurised gas. These microparticles are efficient at delivering DNA into cells, but cannot efficiently transfect small cells and may cause significant tissue damage, thus limiting their potential usefulness. Here we describe the use of 40 nm diameter projectiles - nanoparticles - in biolistic transfections to determine if they are a suitable alternative to microparticles.ResultsExamination of transfection efficiencies in HEK293 cells, using a range of conditions including different DNA concentrations and different preparation procedures, reveals similar behaviour of microparticles and nanoparticles. The use of nanoparticles, however, resulted in ~30% fewer damaged HEK293 cells following transfection. Biolistic transfection of mouse ear tissue revealed similar depth penetration for the two types of particles, and also showed that < 10% of nuclei were damaged in nanoparticle-transfected samples, compared to > 20% in microparticle-transfected samples. Visualising details of small cellular structures was also considerably enhanced when using nanoparticles.ConclusionsWe conclude that nanoparticles are as efficient for biolistic transfection as microparticles, and are more appropriate for use in small cells, when examining cellular structures and/or where tissue damage is a problem.

【 授权许可】

CC BY   
© O'Brien and Lummis; licensee BioMed Central Ltd. 2011