Rat liver-targeted naked plasmid DNA transfer by tail vein injection.
High levels of foreign gene expression in mouse hepatocytes can be achieved by "hydrodynamics-based transfection," the rapid injection of a large volume of a naked deoxyribonucleic acid (DNA) solution into the tail vein. Rats are more tolerant of the frequent phlebotomies required for monitoring blood parameters than mice and, thus, are more suitable for some biomedical research. Recently, we demonstrated that hydrodynamics-based transfection can also be used to deliver naked plasmid DNA into the normal rat, which is more than 10 times larger than the mouse. We performed the tail vein injection using a syringe with a winged needle equipped with an external tube. Injection of a lac Z expression plasmid, pCAGGS-lac Z by this technique resulted in the exclusive detection of beta-galactosidase in the liver. We also injected a rat erythropoietin (Epo) expression plasmid, pCAGGS-Epo (800 microg). Maximal Epo gene expression was achieved when a 25-mL injection volume (approx 100 mL/kg body wt) was transferred within 15 s.
Maruyama
Hiroki
H
Hiroki Maruyama, MD, Division of Clinical Nephrology and Rheumatology, Niigata University Graduate School of Medical and Dental Sciences, 1-754 Asahimachi-dori, Niigata 951-8120, Japan. hirokim@med.niigata-u.ac.jp
Higuchi
Noboru
N
Kameda
Shigemi
S
Miyazaki
Jun-Ichi
J
Gejyo
Fumitake
F
D000465:Algorithms; D000818:Animals; D018390:Gene Targeting / Q000379:methods*; D018014:Gene Transfer Techniques; D007275:Injections, Intravenous / Q000379:methods*; D008099:Liver / Q000378:metabolism*; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar; D013048:Specimen Handling / Q000379:methods; D013623:Tail / Q000098:blood supply