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Cited by 454

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Systematic Determination of the Packaging Limit of Lentiviral Vectors

Kumar, Mukesh, Keller, Brian, Makalou, Ndeye, Sutton, Richard E.·2001·Human Gene Therapy

Abstract

Because of their ability to transduce nondividing cells, human immunodeficiency virus type 1 (HIV)-based vectors have great potential for the therapeutic delivery of genes to cells. We describe here a systematic study of the packaging limit of HIV-based vectors. Restriction endonuclease-generated bacterial chromosomal DNA fragments of different lengths were cloned at three different positions within a lentiviral vector. Vesicular stomatitis virus G protein (VSV G) pseudotyped lentiviral particles were prepared and the different clones were titered on mammalian cells. We observed that the restriction endonuclease site positions at the 5' and 3' ends of the genome were superior with regard to insertional capacity of foreign DNA. In all cases, viral titers decreased semi-logarithmically with increasing vector length. There appears to be no absolute packaging limit because measurable titers were obtained even when the proviral length was in excess of 18 kb. The reduction in titer appears to occur at the level of viral encapsidation, although we cannot exclude limitations in nuclear export of proviral RNA. These results suggest that HIV-based vectors may have a secondary advantage over oncoretroviral vectors because of their greater packaging limit, although the very low titers of the larger vectors will be of limited utility.

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Metadata

Authors
Kumar, M., Keller, B., Makalou, N., & Sutton, R. E.
Venue
Human Gene Therapy
Published
2001-10-10
Volume
12
Issue
15
Pages
1893-1905
ISSN
  • 1043-0342 (ISSN-L)
  • 1557-7422
OpenAlex
W2050870581
Cited by
454
References
18
FWCI
2.93
Type
journal article
Language
en
Open access
closed

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CoChat. (2026). Citation check: Systematic Determination of the Packaging Limit of Lentiviral Vectors. https://app.cochat.ai/papers/10.1089/104303401753153947
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Kumar, M., Keller, B., Makalou, N., & Sutton, R. E. (2001). Systematic Determination of the Packaging Limit of Lentiviral Vectors. Human Gene Therapy. https://doi.org/10.1089/104303401753153947

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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).

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