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LENTI-ONE™ TRANS: TRANSIENT LENTIVIRAL VECTORS

 

  What are Lenti-ONE™ Trans?

Lenti-ONE™ Trans are derived from HIV-1 and are deficient for replication and reverse transcription. The RNA vector genome is thus taken in charge by the host cell machinery as an mRNA and directly translated into a protein. Consequently, Lenti-ONE™ Trans does not involve any DNA form of the genome and are used to mediate fully transient expression in dividing and non-dividing cells. The reverse transcription blockade results from one or two mutations of the reverse transcriptase which is necessary to translate the RNA genomes into DNA. The mutations of the reverse transcriptase that we used completely block the reverse transcription step, consequently, no DNA and residual integration is observed.

Their tropism is defined by the envelop glycoprotein they have been produced with. GEG Tech offers Lenti-ONE™ Trans pseudotyped with VSV glycoprotein, conferring a broad tropism, and Lenti-ONE™ Trans pseudotyped with Mokola glycoprotein, conferring a glial specific tropism. Depending on your purpose and target cells, Lenti-ONE™ Trans are available at High Titer (107 TU/mL) and High Titer+ (108 TU/mL).

 

  What is the difference between the three types of Lenti-ONE™ Trans?

GEG Tech provides three different Lenti-ONE™ Trans which correspond to three different reverse transcriptase mutants: D110E, R178Q and the double mutant D110E/R178Q. Through our experimental results, we didn't point out a significant difference of efficiency or background of reverse transcriptase activity between these three vectors. However, we cannot be sure it will be the case in every experimental context. For this reason we provide these three different Lenti-ONE™ Trans and also a Starter Set of vector which allows to compare them in a particular experimental context.

 

  Why is the expression of Lenti-ONE™ Trans transient in dividing and

non-dividing transduced cells?

The genome of a Lenti-ONE™ Trans remains in an RNA form and is taken care of by the host cell machinery as an endogenous mRNA to be translated into a protein. Consequently, Lenti-ONE™ Trans expression is fully transient in dividing and non-dividing cells. The level of expression is lower than with Lenti-ONE™ or Lenti-ONE™ Epi.

 

  Which cell type is transduced by Lenti-ONE™ Trans?

Unlike Lenti-ONE™ or Lenti-ONE™ Epi, Lenti-ONE™ Trans have no internal promoter. Hence the degree of specificity is only conferred by the envelope. If the envelope is pantropic (e.g. VSV), the expression will be ubiquitous while it can be targeted into specific cell types when using restricting envelope (e.g. Mokola to target glial cells).

 

  Why should I use Lenti-ONE™ Trans?

Lenti-ONE™ Trans do not involve DNA forms of the vector genome, the RNA genome is taken care of as an mRNA of the transduced cells and directly translated into a protein. Hence, Lenti-ONE™ Trans mediate fully transient transgene expression (residual integration is close to 0.1% of that observed with an Lenti-ONE™ Epi). The level of transgene expression is also reduced compared to Lenti-ONE™ and Lenti-ONE™ Epi. Consequently, Lenti-ONE™ Trans are particularly suited for transient expression when high caution is given to the risk of long term expression and the level of expression required is low, for instance:

  • Expression of genome editing tools
  • Expression of differentiation factors