HOMEPRODUCTSCOMPANYCONTACTFAQResearchDictionaryPharmaSign Up FREE or Login

Zinc finger endonuclease targeting PSIP1 inhibits HIV-1 integration.

Abstract
Genome editing using zinc finger nucleases (ZFNs) has been successfully applied to disrupt CCR5 or CXCR4 host factors and inhibit viral entry and infection. Gene therapy using ZFNs to modify the PSIP1 gene, which encodes the lens epithelium-derived growth factor (LEDGF) protein, might restrain an early step of the viral replication cycle at the integration level. ZFNs targeting the PSIP1 gene (ZFNLEDGF) were designed to specifically recognize the sequence after the integrase binding domain (IBD) of the LEDGF/p75 protein. ZFNLEDGF successfully recognized the target region of the PSIP1 gene in TZM-bl cells by heteroduplex formation and DNA sequence analysis. Gene editing induced a frameshift of the coding region and resulted in the abolishment of LEDGF expression at the mRNA and protein levels. Functional assays revealed that infection with the HIV-1 R5 BaL or X4 NL4-3 viral strains was impaired in LEDGF/p75 knockout cells regardless of entry tropism due to a blockade in HIV-1 proviral integration into the host genome. However, residual infection was detected in the LEDGF knockout cells. Indeed, LEDGF knockout restriction was overcome at a high multiplicity of infection, suggesting alternative mechanisms for HIV-1 genome integration rather than through LEDGF/p75. However, the observed residual integration was sensitive to the integrase inhibitor raltegravir. These results demonstrate that the described ZFNLEDGF effectively targets the PSIP1 gene, which is involved in the early steps of the viral replication cycle; thus, ZFNLEDGF may become a potential antiviral agent for restricting HIV-1 integration. Moreover, LEDGF knockout cells represent a potent tool for elucidating the role of HIV integration cofactors in virus replication.
AuthorsRoger Badia, Eduardo Pauls, Eva Riveira-Munoz, Bonaventura Clotet, José A Esté, Ester Ballana
JournalAntimicrobial agents and chemotherapy (Antimicrob Agents Chemother) Vol. 58 Issue 8 Pg. 4318-27 (Aug 2014) ISSN: 1098-6596 [Electronic] United States
PMID24820090 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCopyright © 2014, American Society for Microbiology. All Rights Reserved.
Chemical References
  • Adaptor Proteins, Signal Transducing
  • Anti-HIV Agents
  • PSIP1 protein, human
  • Pyrrolidinones
  • Transcription Factors
  • Raltegravir Potassium
  • HIV Integrase
  • Endonucleases
  • p31 integrase protein, Human immunodeficiency virus 1
Topics
  • Adaptor Proteins, Signal Transducing (antagonists & inhibitors, genetics, metabolism)
  • Amino Acid Sequence
  • Anti-HIV Agents (pharmacology)
  • Endonucleases (genetics, metabolism)
  • Gene Expression Regulation
  • HIV Integrase (genetics, metabolism)
  • HIV-1 (drug effects, genetics)
  • HeLa Cells
  • Host-Pathogen Interactions
  • Humans
  • K562 Cells
  • Molecular Sequence Data
  • Molecular Targeted Therapy
  • Open Reading Frames
  • Plasmids (chemistry, metabolism)
  • Protein Engineering
  • Pyrrolidinones (pharmacology)
  • Raltegravir Potassium
  • Signal Transduction
  • Transcription Factors (antagonists & inhibitors, genetics, metabolism)
  • Transfection
  • Virus Integration (drug effects)
  • Virus Replication (drug effects)
  • Zinc Fingers (genetics)

Join CureHunter, for free Research Interface BASIC access!

Take advantage of free CureHunter research engine access to explore the best drug and treatment options for any disease. Find out why thousands of doctors, pharma researchers and patient activists around the world use CureHunter every day.
Realize the full power of the drug-disease research graph!


Choose Username:
Email:
Password:
Verify Password:
Enter Code Shown: