Optimizing hyaluronidase dose and plasmid DNA delivery greatly improves gene electrotransfer efficiency in rat skeletal muscle

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Dokumenter

Thorbjörn Åkerström, Kenneth Vedel, Josefine Needham Andersen, Pernille Hojman, Eftychia Kontou, Ylva Hellsten, Jørgen Wojtaszewski

Transfection of rat skeletal muscle in vivo is a widely used research model. However, gene electrotransfer protocols have been developed for mice and yield variable results in rats. We investigated whether changes in hyaluronidase pre-treatment and plasmid DNA delivery can improve transfection efficiency in rat skeletal muscle. We found that pre-treating the muscle with a hyaluronidase dose suitable for rats (0.56. U/g b.w.) prior to plasmid DNA injection increased transfection efficiency by >200% whereas timing of the pre-treatment did not affect efficiency. Uniformly distributing plasmid DNA delivery across the muscle by increasing the number of plasmid DNA injections further enhanced transfection efficiency whereas increasing plasmid dose from 0.2 to 1.6. μg/g b.w. or vehicle volume had no effect. The optimized protocol resulted in ~80% (CI95%: 79-84%) transfected muscle fibers with a homogenous distribution. We also show that transfection was stable over five weeks of regular exercise or inactivity. Our findings show that species-specific plasmid DNA delivery and hyaluronidase pre-treatment greatly improves transfection efficiency in rat skeletal muscle.

OriginalsprogEngelsk
TidsskriftBiochemistry and Biophysics Reports
Vol/bind4
Sider (fra-til)342-350
Antal sider9
ISSN2405-5808
DOI
StatusUdgivet - dec. 2015

Bibliografisk note

CURIS 2016 NEXS 274
(Referencen er fra december 2015 men først fundet og registreret i CURIS den 30. september 2016!)

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