Production of Viral Vectors with Suicide Genes by Utilizing the Intron-Splicing Mechanism of Insect Cells

ABSTRACT This report describes novel baculovirus vectors designed to express mammalian β1,4-galactosyltransferase and α2,6-sialyltransferase genes at early times after infection. Sf9 cells infected with these viral vectors, unlike cells infected with a wild-type baculovirus, produced a sialylated viral glycoprotein during the late phase of infection. Thus, the two mammalian glycosyltransferases encoded by these viral vectors are necessary and sufficient for sialylation of a foreign glycoprotein in insect cells under the conditions used in this study. While some of the new baculovirus vectors described in this study produced less, one produced wild-type levels of infectious budded virus progeny.

Download Full-text

Abstract LB-368: Exploiting the mechanism of intron-splicing to produce viral vectors harboring toxin genes for cancer therapy

10.1158/1538-7445.am2012-lb-368 ◽

2012 ◽

Author(s):

Haifeng Chen

Keyword(s):

Cancer Therapy ◽

Viral Vectors ◽

Intron Splicing ◽

Toxin Genes

Download Full-text

Production of adeno-associated viral vectors in insect cells using triple infection: Optimization of baculovirus concentration ratios

Biotechnology and Bioengineering ◽

10.1002/bit.21069 ◽

2006 ◽

Vol 95 (6) ◽

pp. 1081-1092 ◽

Cited By ~ 34

Author(s):

Marc G. Aucoin ◽

Michel Perrier ◽

Amine A. Kamen

Keyword(s):

Viral Vectors ◽

Insect Cells ◽

Triple Infection ◽

Concentration Ratios

Download Full-text

Acquisition of Complement Resistance through Incorporation of CD55/Decay-Accelerating Factor into Viral Particles Bearing Baculovirus GP64

Journal of Virology ◽

10.1128/jvi.02519-09 ◽

2010 ◽

Vol 84 (7) ◽

pp. 3210-3219 ◽

Cited By ~ 29

Author(s):

Yuuki Kaname ◽

Hideki Tani ◽

Chikako Kataoka ◽

Mai Shiokawa ◽

Shuhei Taguwa ◽

...

Keyword(s):

Envelope Protein ◽

Viral Vectors ◽

Molecular Mechanisms ◽

Insect Cells ◽

Decay Accelerating Factor ◽

Complement Regulatory Proteins ◽

Complement Resistance ◽

Viral Particles ◽

Lentivirus Vectors

ABSTRACT A major obstacle to gene transduction by viral vectors is inactivation by human complement in vivo. One way to overcome this is to incorporate complement regulatory proteins, such as CD55/decay accelerating factor (DAF), into viral particles. Lentivirus vectors pseudotyped with the baculovirus envelope protein GP64 have been shown to acquire more potent resistance to serum inactivation and longer transgene expression than those pseudotyped with the vesicular stomatitis virus (VSV) envelope protein G. However, the molecular mechanisms underlying resistance to serum inactivation in pseudotype particles bearing the GP64 have not been precisely elucidated. In this study, we generated pseudotype and recombinant VSVs bearing the GP64. Recombinant VSVs generated in human cell lines exhibited the incorporation of human DAF in viral particles and were resistant to serum inactivation, whereas those generated in insect cells exhibited no incorporation of human DAF and were sensitive to complement inactivation. The GP64 and human DAF were detected on the detergent-resistant membrane and were coprecipitated by immunoprecipitation analysis. A pseudotype VSV bearing GP64 produced in human DAF knockdown cells reduced resistance to serum inactivation. In contrast, recombinant baculoviruses generated in insect cells expressing human DAF or carrying the human DAF gene exhibited resistance to complement inactivation. These results suggest that the incorporation of human DAF into viral particles by interacting with baculovirus GP64 is involved in the acquisition of resistance to serum inactivation.

Download Full-text