9. When aspirating and replacing medium, aspirate three wells at a
time and immediately replace the medium with serum-free
medium. This approach minimizes the time the cells are
allowed to dry, which can lead to the cells detaching from the
plate.
10. The protocol describes transduction of a CAR-negative cell
line, NIH/3T3. Depending on the particular cell line used, it
may be necessary to vary the cell culture medium, cell seeding
density, and MOI.
Acknowledgments
This work was supported in part by an NSF CAREER Award
(DMR 1352535).
References
1. Ginn SL et al (2018) Gene therapy clinical trials
worldwide to 2017: an update. J Gene Med 20
(5):e3015
2. Tatsis N, Ertl HC (2004) Adenoviruses as vaccine vectors. Mol Ther 10(4):616–629
3. Wohlfart C (1988) Neutralization of adenoviruses: kinetics, stoichiometry, and mechanisms. J Virol 62(7):2321–2328
4. Nayak S, Herzog RW (2010) Progress and
prospects: immune responses to viral vectors.
Gene Ther 17(3):295
5. Worgall S et al (1997) Innate immune mechanisms dominate elimination of adenoviral vectors following in vivo administration. Hum
Gene Ther 8(1):37–44
6. Sharma A et al (2009) Adenovirus receptors
and their implications in gene delivery. Virus
Res 143(2):184–194
7. Bergelson JM et al (1997) Isolation of a common receptor for Coxsackie B viruses and adenoviruses
2
and
5.
Science
275
(5304):1320–1323
8. Wu H et al (2002) Construction and characterization of adenovirus serotype 5 packaged by
serotype
3
hexon.
J
Virol
76
(24):12775–12782
9. Volk AL et al (2003) Enhanced adenovirus
infection of melanoma cells by fibermodification: incorporation of RGD peptide
or Ad5/3 chimerism. Cancer Biol Ther 2
(5):511–515
10. Fisher K et al (2001) Polymer-coated adenovirus permits efficient retargeting and evades
neutralising antibodies. Gene Ther 8(5):341
11. O’Riordan C, Lachapelle A, Delgado C, Parkes
V (1999) Wadsworth SC Smith AE Francis GE
PEGylation of adenovirus with retention of
infectivity and protection from neutralizing
antibody in vitro and in vivo. Hum Gene Ther
10:1349–1358
12. Baker A et al (1997) Polyethylenimine (PEI) is
a simple, inexpensive and effective reagent for
condensing and linking plasmid DNA to adenovirus for gene delivery. Gene Ther 4(8):773
13. Kawamata Y et al (2002) Receptorindependent augmentation of adenovirusmediated gene transfer with chitosan in vitro.
Biomaterials 23(23):4573–4579
14. Fasbender A et al (1997) Complexes of adenovirus with polycationic polymers and cationic
lipids increase the efficiency of gene transfer
in vitro and in vivo. J Biol Chem 272
(10):6479–6489
15. Croyle MA et al (2001) “Stealth” adenoviruses
blunt cell-mediated and humoral immune
responses against the virus and allow for significant gene expression upon readministration in
the lung. J Virol 75(10):4792–4801
16. Wonganan P, Croyle MA (2010) PEGylated
adenoviruses: from mice to monkeys. Viruses
2(2):468–502
17. Kanerva A, Hemminki A (2004) Modified adenoviruses for cancer gene therapy. Int J Cancer
110(4):475–480
18. Dmitriev I et al (1998) An adenovirus vector
with genetically modified fibers demonstrates
expanded tropism via utilization of a coxsackievirus and adenovirus receptor-independent
Surface Modification of Adenovirus Vector
365
time and immediately replace the medium with serum-free
medium. This approach minimizes the time the cells are
allowed to dry, which can lead to the cells detaching from the
plate.
10. The protocol describes transduction of a CAR-negative cell
line, NIH/3T3. Depending on the particular cell line used, it
may be necessary to vary the cell culture medium, cell seeding
density, and MOI.
Acknowledgments
This work was supported in part by an NSF CAREER Award
(DMR 1352535).
References
1. Ginn SL et al (2018) Gene therapy clinical trials
worldwide to 2017: an update. J Gene Med 20
(5):e3015
2. Tatsis N, Ertl HC (2004) Adenoviruses as vaccine vectors. Mol Ther 10(4):616–629
3. Wohlfart C (1988) Neutralization of adenoviruses: kinetics, stoichiometry, and mechanisms. J Virol 62(7):2321–2328
4. Nayak S, Herzog RW (2010) Progress and
prospects: immune responses to viral vectors.
Gene Ther 17(3):295
5. Worgall S et al (1997) Innate immune mechanisms dominate elimination of adenoviral vectors following in vivo administration. Hum
Gene Ther 8(1):37–44
6. Sharma A et al (2009) Adenovirus receptors
and their implications in gene delivery. Virus
Res 143(2):184–194
7. Bergelson JM et al (1997) Isolation of a common receptor for Coxsackie B viruses and adenoviruses
2
and
5.
Science
275
(5304):1320–1323
8. Wu H et al (2002) Construction and characterization of adenovirus serotype 5 packaged by
serotype
3
hexon.
J
Virol
76
(24):12775–12782
9. Volk AL et al (2003) Enhanced adenovirus
infection of melanoma cells by fibermodification: incorporation of RGD peptide
or Ad5/3 chimerism. Cancer Biol Ther 2
(5):511–515
10. Fisher K et al (2001) Polymer-coated adenovirus permits efficient retargeting and evades
neutralising antibodies. Gene Ther 8(5):341
11. O’Riordan C, Lachapelle A, Delgado C, Parkes
V (1999) Wadsworth SC Smith AE Francis GE
PEGylation of adenovirus with retention of
infectivity and protection from neutralizing
antibody in vitro and in vivo. Hum Gene Ther
10:1349–1358
12. Baker A et al (1997) Polyethylenimine (PEI) is
a simple, inexpensive and effective reagent for
condensing and linking plasmid DNA to adenovirus for gene delivery. Gene Ther 4(8):773
13. Kawamata Y et al (2002) Receptorindependent augmentation of adenovirusmediated gene transfer with chitosan in vitro.
Biomaterials 23(23):4573–4579
14. Fasbender A et al (1997) Complexes of adenovirus with polycationic polymers and cationic
lipids increase the efficiency of gene transfer
in vitro and in vivo. J Biol Chem 272
(10):6479–6489
15. Croyle MA et al (2001) “Stealth” adenoviruses
blunt cell-mediated and humoral immune
responses against the virus and allow for significant gene expression upon readministration in
the lung. J Virol 75(10):4792–4801
16. Wonganan P, Croyle MA (2010) PEGylated
adenoviruses: from mice to monkeys. Viruses
2(2):468–502
17. Kanerva A, Hemminki A (2004) Modified adenoviruses for cancer gene therapy. Int J Cancer
110(4):475–480
18. Dmitriev I et al (1998) An adenovirus vector
with genetically modified fibers demonstrates
expanded tropism via utilization of a coxsackievirus and adenovirus receptor-independent
Surface Modification of Adenovirus Vector
365