Lentivirus-Retrovirus

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What Are Lentiviruses and Retroviruses?

Lentiviruses and retroviruses are RNA viruses from the Retroviridae family, widely used for stable gene delivery in dividing and non-dividing cells. These vectors integrate into the host genome, allowing for long-term gene expression and can package target sequences up 8 kb (total size between the LTRs).

  • Lentiviruses (e.g., HIV-1-based vectors) can transduce a wide range of dividing and non-dividing cells, including neurons and stem cells.
  • Retroviruses (e.g., MMLV-based vectors) efficiently transduce only dividing cells.

Pseudotyping

Pseudotyping refers to the process of packaging a viral genome with an envelope protein from another virus, which alters the host range and tropism of the vector. For example, lentiviruses and retroviruses are commonly pseudotyped with the VSV-G envelope to enable broad infectivity across many cell types. Other envelope proteins can be used for more specific targeting.

Retroviral host range depends on the envelope used:

  • Ecotropic retroviruses (infect rodent cells only) → BSL-1
  • Amphotropic retroviruses (infect human and rodent cells) → BSL-2

Lentiviral vectors are commonly pseudotyped with VSV-G and are always handled at BSL-2.

Detailed information and guides about Lentiviral or Retroviral vector systems can be found on Addgene.

Production

Lentiviral or retroviral particles are produced via transient plasmid transfection in LentiX-293T cells. You can choose between:

  • Crude preps (suitable for most standard applications, e.g. generation of stable cell lines)
  • Concentrated virus (via ultracentrifugation for higher titer needs)

Functional titers of concentrated viral preps are determined via FACS or by Limited Dilution Colony Counting depending on presence of fluorescence or selective marker.

The Virus Service @ MBS, ISTA offers custom Lenti- and Retrovirus production and consultation services upon request.