An adenoviral vector for probing promoter activity in primary immune cells. Academic Article uri icon

Overview

abstract

  • Functional analysis of the DNA regulatory regions that control gene expression has largely been performed through transient transfection of promoter-reporter constructs into transformed cells. However, transformed cells are often poor models of primary cells. To directly analyze DNA regulatory regions in primary cells, we generated a novel adenoviral luciferase reporter vector, pShuttle-luciferase-GFP (pSLUG) that contains a promoterless luciferase cassette (with an upstream cloning site) for probing promoter activity, and a GFP expression cassette that allows for the identification of transduced cells. Recombinant adenoviruses generated from this vector can transduce a wide range of primary immune cells with high efficiency, including human macrophages, dendritic cells and T cells; and mouse T cells transgenic for the coxsackie and adenoviral receptor (CAR). In primary T cells, we show inducible nuclear factor of activated T cells (NF-AT) activity using a recombinant pSLUG adenovirus containing a consensus NF-AT promoter. We further show inducible IL-12/23 p40 promoter activity in primary macrophages and dendritic cells using a recombinant pSLUG adenovirus containing the proximal human IL-12/23 p40 promoter. The pSLUG system promises to be a powerful tool for the analysis of DNA regulatory regions in diverse types of primary immune cells.

publication date

  • February 20, 2006

Research

keywords

  • Adenoviridae
  • Genetic Vectors
  • Promoter Regions, Genetic
  • Regulatory Sequences, Nucleic Acid
  • T-Lymphocytes
  • Transduction, Genetic

Identity

PubMed Central ID

  • PMC2964867

Scopus Document Identifier

  • 33646093262

Digital Object Identifier (DOI)

  • 10.1016/j.jim.2006.01.009

PubMed ID

  • 16563424

Additional Document Info

volume

  • 311

issue

  • 1-2