TY - JOUR
T1 - Optimizing hyaluronidase dose and plasmid DNA delivery greatly improves gene electrotransfer efficiency in rat skeletal muscle
AU - Åkerström, Thorbjörn
AU - Vedel, Kenneth
AU - Needham Andersen, Josefine
AU - Hojman, Pernille
AU - Kontou, Eftychia
AU - Hellsten, Ylva
AU - Wojtaszewski, Jørgen
N1 - CURIS 2016 NEXS 274
(Referencen er fra december 2015 men først fundet og registreret i CURIS den 30. september 2016!)
PY - 2015/12
Y1 - 2015/12
N2 - 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.
AB - 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.
KW - Exercise
KW - Gene electrotransfer
KW - Hyaluronidase
KW - Rat
KW - Skeletal muscle
UR - http://www.scopus.com/inward/record.url?scp=84946398527&partnerID=8YFLogxK
U2 - 10.1016/j.bbrep.2015.10.007
DO - 10.1016/j.bbrep.2015.10.007
M3 - Journal article
AN - SCOPUS:84946398527
SN - 2405-5808
VL - 4
SP - 342
EP - 350
JO - Biochemistry and Biophysics Reports
JF - Biochemistry and Biophysics Reports
ER -