Optimizing hyaluronidase dose and plasmid DNA delivery greatly improves gene electrotransfer efficiency in rat skeletal muscle
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Dokumenter
- Akerstrom et al_Biochemistry and Biophysics Reports_2015_Vol 4_342-350
Forlagets udgivne version, 2,83 MB, PDF-dokument
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.
Originalsprog | Engelsk |
---|---|
Tidsskrift | Biochemistry and Biophysics Reports |
Vol/bind | 4 |
Sider (fra-til) | 342-350 |
Antal sider | 9 |
ISSN | 2405-5808 |
DOI | |
Status | Udgivet - dec. 2015 |
Bibliografisk note
CURIS 2016 NEXS 274
(Referencen er fra december 2015 men først fundet og registreret i CURIS den 30. september 2016!)
Antal downloads er baseret på statistik fra Google Scholar og www.ku.dk
ID: 166322375