Production of high-titer lentiviral particles for stable genetic modification of mammalian cells

Michael R. Larcombe, Jan Manent, Joseph Chen, Ketan Mishra, Xiaodong Liu, Christian M. Nefzger

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

2 Citations (Scopus)

Abstract

Lentiviral gene transfer technologies exploit the natural efficiency of viral transduction to integrate exogenous genes into mammalian cells. This provides a simple research tool for inducing transgene expression or endogenous gene knockdown in both dividing and nondividing cells. This chapter describes an improved protocol for polyethylenimine (PEI)-mediated multi-plasmid transfection and polyethylene glycol (PEG) precipitation to generate and concentrate lentiviral vectors.

Original languageEnglish
Title of host publicationMouse Cell Culture
Subtitle of host publicationMethods and Protocols
EditorsIvan Bertoncello
Place of PublicationNew York
PublisherHumana Press
Pages47-61
Number of pages15
ISBN (Electronic)9781493990863
ISBN (Print)9781493990856
DOIs
Publication statusPublished - 1 Jan 2019

Publication series

NameMethods in Molecular Biology
Volume1940
ISSN (Print)1064-3745

Keywords

  • Gene transfer
  • PEG precipitation
  • PEI transfection
  • Titration

Cite this

Larcombe, M. R., Manent, J., Chen, J., Mishra, K., Liu, X., & Nefzger, C. M. (2019). Production of high-titer lentiviral particles for stable genetic modification of mammalian cells. In I. Bertoncello (Ed.), Mouse Cell Culture: Methods and Protocols (pp. 47-61). (Methods in Molecular Biology; Vol. 1940). New York: Humana Press. https://doi.org/10.1007/978-1-4939-9086-3_4
Larcombe, Michael R. ; Manent, Jan ; Chen, Joseph ; Mishra, Ketan ; Liu, Xiaodong ; Nefzger, Christian M. / Production of high-titer lentiviral particles for stable genetic modification of mammalian cells. Mouse Cell Culture: Methods and Protocols. editor / Ivan Bertoncello. New York : Humana Press, 2019. pp. 47-61 (Methods in Molecular Biology).
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Larcombe, MR, Manent, J, Chen, J, Mishra, K, Liu, X & Nefzger, CM 2019, Production of high-titer lentiviral particles for stable genetic modification of mammalian cells. in I Bertoncello (ed.), Mouse Cell Culture: Methods and Protocols. Methods in Molecular Biology, vol. 1940, Humana Press, New York, pp. 47-61. https://doi.org/10.1007/978-1-4939-9086-3_4

Production of high-titer lentiviral particles for stable genetic modification of mammalian cells. / Larcombe, Michael R.; Manent, Jan; Chen, Joseph; Mishra, Ketan; Liu, Xiaodong; Nefzger, Christian M.

Mouse Cell Culture: Methods and Protocols. ed. / Ivan Bertoncello. New York : Humana Press, 2019. p. 47-61 (Methods in Molecular Biology; Vol. 1940).

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

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Larcombe MR, Manent J, Chen J, Mishra K, Liu X, Nefzger CM. Production of high-titer lentiviral particles for stable genetic modification of mammalian cells. In Bertoncello I, editor, Mouse Cell Culture: Methods and Protocols. New York: Humana Press. 2019. p. 47-61. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9086-3_4