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

VL - 4

SP - 342

EP - 350

JO - Biochemistry and Biophysics Reports

JF - Biochemistry and Biophysics Reports

SN - 2405-5808

ER -