Scripps VIVO scripps research logo

  • Index
  • Log in
  • Home
  • People
  • Organizations
  • Research
  • Events
Search form

Zinc finger protein designed to target 2-long terminal repeat junctions interferes with human immunodeficiency virus integration

Academic Article
uri icon
  • Overview
  • Identity
  • Additional Document Info
  • View All
scroll to property group menus

Overview

authors

  • Sakkhachornphop, S.
  • Barbas III, Carlos
  • Keawvichit, R.
  • Wongworapat, K.
  • Tayapiwatana, C.

publication date

  • September 2012

journal

  • Human Gene Therapy  Journal

abstract

  • Integration of the human immunodeficiency virus type 1 (HIV-1) genome into the host chromosome is a vital step in the HIV life cycle. The highly conserved cytosine-adenine (CA) dinucleotide sequence immediately upstream of the cleavage site is crucial for integrase (IN) activity. As this viral enzyme has an important role early in the HIV-1 replication cycle, interference with the IN substrate has become an attractive strategy for therapeutic intervention. We demonstrated that a designed zinc finger protein (ZFP) fused to green fluorescent protein (GFP) targets the 2-long terminal repeat (2-LTR) circle junctions of HIV-1 DNA with nanomolar affinity. We report now that 2LTRZFP-GFP stably transduced into 293T cells interfered with the expression of vesicular stomatitis virus glycoprotein (VSV-G)-pseudotyped lentiviral red fluorescent protein (RFP), as shown by the suppression of RFP expression. We also used a third-generation lentiviral vector and pCEP4 expression vector to deliver the 2LTRZFP-GFP transgene into human T-lymphocytic cells, and a stable cell line for long-term expression studies was selected for HIV-1 challenge. HIV-1 integration and replication were inhibited as measured by Alu-gag real-time PCR and p24 antigen assay. In addition, the molecular activity of 2LTRZFP-GFP was evaluated in peripheral blood mononuclear cells. The results were confirmed by Alu-gag real-time PCR for integration interference. We suggest that the expression of 2LTRZFP-GFP limited viral integration on intracellular immunization, and that it has potential for use in HIV gene therapy in the future.

subject areas

  • Cell Line
  • DNA-Binding Proteins
  • Genetic Therapy
  • Green Fluorescent Proteins
  • HIV Infections
  • HIV Long Terminal Repeat
  • HIV-1
  • Humans
  • T-Lymphocytes
  • Transduction, Genetic
  • Virus Integration
  • Zinc Fingers
scroll to property group menus

Identity

PubMed Central ID

  • PMC3440019

International Standard Serial Number (ISSN)

  • 1043-0342

Digital Object Identifier (DOI)

  • 10.1089/hum.2011.124

PubMed ID

  • 22429108
scroll to property group menus

Additional Document Info

start page

  • 932

end page

  • 942

volume

  • 23

issue

  • 9

©2021 The Scripps Research Institute | Terms of Use | Powered by VIVO

  • About
  • Contact Us
  • Support