Improvement in adenoviral gene transfer efficiency after preincubation at +37 degrees C in vitro and in vivo.
Adenovirus is a widely used vector in gene transfer experiments because it produces high transduction efficiency in vitro and in vivo by means of the coxsackie-adenovirus receptor (CAR) and major histocompatibility complex (MHC) class I alpha-2 domain. Adenoviral gene transfer efficiency has been reported to correlate with cellular CAR expression. We report here a simple method to increase adenoviral gene transfer efficiency in cells that do not express high levels of CAR: preincubation of adenovirus for 30-40 minutes at +37 degrees C significantly increased the transduction efficiency in vitro in CHO and BALB/3T3 cells, in which CAR is expressed at very low levels. Increased transduction efficiency of preincubated adenovirus was also detected in vivo in rat brain tissue. In addition, we found that adenoviruses were rapidly inactivated in human serum in a complement-independent manner, whereas fetal bovine serum (FBS) had hardly any effects on the viral infectivity. We conclude that preincubation of adenoviral vectors at +37 degrees C may substantially increase gene transfer efficiency in applications in which target cells do not express high levels of CAR.
Kossila
Maija
M
A. I. Virtanen Institute for Molecular Sciences, University of Kuopio, Kuopio, Finland FIN-70211.
Jauhiainen
Suvi
S
Laukkanen
Mikko O
MO
Lehtolainen
Pauliina
P
Jääskeläinen
Maiju
M
Turunen
Päivi
P
Loimas
Sami
S
Wahlfors
Jarmo
J
Ylä-Herttuala
Seppo
S
D016475:3T3 Cells; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D006358:Hot Temperature; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D014161:Transduction, Genetic
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins