Protocols

INTEGRATING AND NON-INTEGRATING LENTIVIRAL VECTORS

Lentiviral vectors are very efficient tools for delivery DNA sequences of interest in vitro and in vivo. Their high titer production and their high flexibility of pseudotyping make Lentiviral vectors a tool of choice in gene transfer. Furthermore, GEG Tech offers multiple mutants, issued from GEG Tech R&D team innovations. The modification of some features of lentiviral vectors enlarges the scope of applications of these tools.

INTEGRATING AND NON-INTEGRATING LENTIVIRAL VECTORS

In their original form, they allow stable integration of their genome in the target cell chromosomes. The transgene expression they mediate is thus maintained in dividing cells.

Integrase-Deficient Lentiviral Vectors

Integration-Deficient lentiviral vectors (mutations D64V, N and LQ) lead to the generation of episomal vector genomes. They can be used for transient expression in dividing cells or long term expression in non-dividing cells without the limits inherent to the integration of the vector.

  •  in vitro and in vivo long term expression in non-dividing cells or transient expression in dividing cells
  •  Genome editing tools delivery for in vitro and in vivo applications and transgenesis (KO/KI)
  •  IPSCs Generation

Integrase-Deficient Lentiviral Vectors

Reverse transcription-Deficient lentiviral vectors result from the mutation of the reverse-transcriptase (mutations D110E, E478Q and D110E/E478Q), which leads to the production of a vector whose genome cannot be reverse-transcribed. Consequently, there is no DNA synthesis and the RNA vector genome is taken in charge by the transduced cell machinery like a mRNA. Those vectors allow a fully transient expression regardless of the type of cells (dividing or not).

  •  Transient expression of therapeutic factors
  •  Genome editing tools delivery for in vitro and in vivo applications and transgenesis (KO/KI)

SCOPE OF APPLICATIONS

Comparison of application possibilities offered by mostly used gene transfer methods

Long Term Expression

Transient Expression

Generation of Stable Cell Lines

Transgenesis

Non-dividing Cells
Dividing Cells
Non-dividing Cells
Dividing Cells
Plasmids
-
-
+
+
+
-
Adenoviral Vectors
+
-
-
+
-
-
Adeno Ass. Viral Vectors
+
-
-
+
-
-
Retroviral Vectors
-
+
-
-
+
+
Lentiviral Vectors
+
+
-
-
+
+
Lenti-ONE Vectors
+
+
+
+
+
+

Long Term Expression

Transient Expression

Generation of Stable Cell Lines

Transgenesis

Non-dividing Cells
Dividing Cells
Non-dividing Cells
Dividing Cells
Plasmids
-
-
+
+
+
-
Adenoviral Vectors
+
-
-
+
-
-
Adeno Ass. Viral Vectors
+
-
-
+
-
-
Retroviral Vectors
-
+
-
-
+
+
Lentiviral Vectors
+
+
-
-
+
+
Lenti-ONE Vectors
+
+
+
+
+
+

LENTIVIRAL VECTORS

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