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MMLV-Based Retrovirus Service

Recombinant MMLV retroviral vectors are derived from Moloney murine leukemia virus and has been widely used for delivering exogenous genes into a wide range of dividing cells. MMLV can integrate into the host genome and has the ability to mediate potent transduction and stable expression of a specific nucleic acid sequence into the target cell's genome both in vitro and in vivo. The viral genome of MMLV is a single stranded, positive-sense RNA containing gag, pol, and env, coding for structural proteins, reverse transcriptase and coat proteins, respectively.

The development of recombinant MMLV retroviruses involves a transfer plasmid, packaging plasmids and a packaging cell line. The transfer plasmid contains a number of cis-acting viral elements such as a viral packaging signal, a transgene or shRNA of interest, which are flanked by long terminal repeat (LTR) sequences. The packaging plasmids contain genes encoding envelop protein and viral proteins for replication and packaging. The virus production is accomplished by co-transfection of these plasmids into a packaging cell line.

Schematic diagram for MMLV retrovirus production Figure 1. Schematic diagram for MMLV retrovirus production

Creative Biogene offers custom services for development and production of MMLV-based, replication-defective and VSV-G pseudotyped retrovirus. Genes required for viral packaging and transduction are designed to be replaced with gene(s) or sequence elements to be transferred. Viral proteins needed for the initial transduction are provided in trans by other plasmids or the packaging cell line. As a result, the produced retroviruses can transduce target cells but can not further replicate in the transduced target cell. The use of the VSV-G envelope glycoprotein from vesicular stomatitis virus, allows a wide host tropism and enable the virus to transduce many cell types.

Applications

-Gene delivery into a wide type of cultured dividing cells
-Stable cell line generation
-In vivo animal tests

Features

-One-stop solution: from vector design, to DNA synthesis, to MMLV-based retrovirus production
-Produce virus in high titer that can be used for in vivo injection
-Rigorous quality controls

References:

  1. Gilboa E, Kolbe M, Noonan K, Kucherlapati R. Construction of a mammalian transducing vector from the genome of Moloney murine leukemia virus. J. Virol. 1982;44:845–851.
  2. Freed EO, Risser R. The role of envelope glycoprotein processing in murine leukemia virus infection. J. Virol. 1987;61:2852–2856.
  3. Miller DG, Adam MA, Miller AD. Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection. Mol. Cell Biol. 1990;10:4239–4242.
For research use only. Not intended for any clinical use.