Abstract
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 1 α-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°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°C may substantially increase gene transfer efficiency in applications in which target cells do not express high levels of CAR.
Original language | English |
---|---|
Pages (from-to) | 87-93 |
Number of pages | 7 |
Journal | Molecular Therapy |
Volume | 5 |
Issue number | 1 |
DOIs | |
Publication status | Published - 2002 |
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Keywords
- Adenovirus
- Coxsackie adenovirus receptor
- Gene transfer
- Serum
ASJC Scopus subject areas
- Molecular Biology
Cite this
Improvement in adenoviral gene transfer efficiency after preincubation at +37°C in vitro and in vivo. / Kossila, Maija; Jauhiainen, Suvi; Laukkanen, Mikko O.; Lehtolainen, Pauliina; Jääskeläinen, Maiju; Turunen, Päivi; Loimas, Sami; Wahlfors, Jarmo; Ylä-Herttuala, Seppo.
In: Molecular Therapy, Vol. 5, No. 1, 2002, p. 87-93.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Improvement in adenoviral gene transfer efficiency after preincubation at +37°C in vitro and in vivo
AU - Kossila, Maija
AU - Jauhiainen, Suvi
AU - Laukkanen, Mikko O.
AU - Lehtolainen, Pauliina
AU - Jääskeläinen, Maiju
AU - Turunen, Päivi
AU - Loimas, Sami
AU - Wahlfors, Jarmo
AU - Ylä-Herttuala, Seppo
PY - 2002
Y1 - 2002
N2 - 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 1 α-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°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°C may substantially increase gene transfer efficiency in applications in which target cells do not express high levels of CAR.
AB - 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 1 α-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°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°C may substantially increase gene transfer efficiency in applications in which target cells do not express high levels of CAR.
KW - Adenovirus
KW - Coxsackie adenovirus receptor
KW - Gene transfer
KW - Serum
UR - http://www.scopus.com/inward/record.url?scp=0036154969&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0036154969&partnerID=8YFLogxK
U2 - 10.1006/mthe.2001.0516
DO - 10.1006/mthe.2001.0516
M3 - Article
C2 - 11786050
AN - SCOPUS:0036154969
VL - 5
SP - 87
EP - 93
JO - Molecular Therapy
JF - Molecular Therapy
SN - 1525-0016
IS - 1
ER -