Improved recombinant retroviral titers utilizing trichostatin A.

Abstract:

:Recombinant retroviruses are a common vehicle to deliver an exogenous gene to a target cell, which makes them a useful tool in the field of gene therapy. A major drawback to using recombinant retroviruses is the low titer achieved, resulting in a limited number of target cells infected and subsequently poor expression of a transgene. In this study, we created an ecotropic producer cell line that contained recombinant mouse nerve growth factor (NGF) and the reporter gene LacZ. This cell line, termed psi 2/LIG/NGF, was treated with the drug trichostatin A, an inhibitor of histone deacetylase. At a concentration of 3 microM trichostatin A, the retroviral titer of this producer cell line was increased 34-fold relative to untreated control cells and 3.4-fold compared to the commonly used hyperacetylating compound sodium butyrate. Producer cells treated with trichostatin A also increased the number of primary rat marrow stromal cells infected 5.8-fold compared to untreated producer cells. These results offer a simple and effective solution for converting low titer producer cell clones to higher titer clones, which can be easily tested and applied.

journal_name

Biotechniques

journal_title

BioTechniques

authors

Tobias CA,Kim D,Fischer I

doi

10.2144/00294rr05

subject

Has Abstract

pub_date

2000-10-01 00:00:00

pages

884-90

issue

4

eissn

0736-6205

issn

1940-9818

journal_volume

29

pub_type

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