Zinc finger protein designed to target 2-long terminal repeat junctions interferes with human immunodeficiency virus integration
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 enzym...
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th-cmuir.6653943832-9092014-08-29T09:02:19Z Zinc finger protein designed to target 2-long terminal repeat junctions interferes with human immunodeficiency virus integration Sakkhachornphop S. Barbas C.F. Keawvichit R. Wongworapat K. Tayapiwatana C. 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. © 2012, Mary Ann Liebert, Inc. 2014-08-29T09:02:19Z 2014-08-29T09:02:19Z 2012 Article 10430342 10.1089/hum.2011.124 HGTHE http://www.scopus.com/inward/record.url?eid=2-s2.0-84866325679&partnerID=40&md5=8566b0689be01855e3a4fb2b8a76e98c http://cmuir.cmu.ac.th/handle/6653943832/909 English |
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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. © 2012, Mary Ann Liebert, Inc. |
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Article |
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Sakkhachornphop S. Barbas C.F. Keawvichit R. Wongworapat K. Tayapiwatana C. |
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Sakkhachornphop S. Barbas C.F. Keawvichit R. Wongworapat K. Tayapiwatana C. Zinc finger protein designed to target 2-long terminal repeat junctions interferes with human immunodeficiency virus integration |
author_facet |
Sakkhachornphop S. Barbas C.F. Keawvichit R. Wongworapat K. Tayapiwatana C. |
author_sort |
Sakkhachornphop S. |
title |
Zinc finger protein designed to target 2-long terminal repeat junctions interferes with human immunodeficiency virus integration |
title_short |
Zinc finger protein designed to target 2-long terminal repeat junctions interferes with human immunodeficiency virus integration |
title_full |
Zinc finger protein designed to target 2-long terminal repeat junctions interferes with human immunodeficiency virus integration |
title_fullStr |
Zinc finger protein designed to target 2-long terminal repeat junctions interferes with human immunodeficiency virus integration |
title_full_unstemmed |
Zinc finger protein designed to target 2-long terminal repeat junctions interferes with human immunodeficiency virus integration |
title_sort |
zinc finger protein designed to target 2-long terminal repeat junctions interferes with human immunodeficiency virus integration |
publishDate |
2014 |
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http://www.scopus.com/inward/record.url?eid=2-s2.0-84866325679&partnerID=40&md5=8566b0689be01855e3a4fb2b8a76e98c http://cmuir.cmu.ac.th/handle/6653943832/909 |
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1681419571794280448 |