# RelA and mitogen-activated protein kinase kinase kinases potently enhance lentiviral vector production

## TL;DR

This study shows that using RelA and MAP3Ks in producer cells can significantly boost the production of lentiviral vectors used in gene therapy.

## Contribution

The paper introduces RelA and MAP3Ks as novel factors to enhance lentiviral vector production in producer cells.

## Key findings

- Co-expression of RelA or MAP3Ks with packaging constructs enhances lentiviral vector production 3- to 4-fold.
- Adding IκB kinase 2 or HIV Tat further increases the production boost from RelA or MAP3Ks.
- RelA and MAP3K1 also enhance lentiviral production in suspension-grown HEK293T/17SF cells.

## Abstract

The growing demands for gene therapy have encouraged development of safe and efficient lentiviral vector (LV) preparation. While much progress has been made in this field, it remains to be explored how to boost its production from producer cells. This paper reports that transient co-expression of RelA or several mitogen-activated protein kinase kinase kinases (MAP3Ks) with packaging constructs can potently enhance LV production in HEK293T producer cells. Adding in transfection a small amount of effector plasmid is sufficient to achieve 3- to 4-fold enhancement, which can further be augmented by co-expression of IκB kinase 2 or HIV Tat. It is also shown that expression of RelA or MAP3K1 can increase LV production in HEK293T/17SF cells grown in suspension. These results indicate that stimulation of intracellular signaling pathways in producer cells represents a powerful means for enhancing LV production.

•RelA increases production of lentiviral vector.•MAPKKKs increase production of lentiviral vector.•Tat further enhances lentiviral production by RelA or MAPKKKs.•RelA and MAPKKK1 increase lentiviral production in suspension cells.

RelA increases production of lentiviral vector.

MAPKKKs increase production of lentiviral vector.

Tat further enhances lentiviral production by RelA or MAPKKKs.

RelA and MAPKKK1 increase lentiviral production in suspension cells.