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2ltrzfp interacts specifically to hiv-1 dna without off-target effects as determined by biolayer interferometry
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Document Title
2ltrzfp interacts specifically to hiv-1 dna without off-target effects as determined by biolayer interferometry
Author
Chupradit K., Thongkum W., Juntit O.-A., Sornsuwan K., Tayapiwatana C.
Name from Authors Collection
Affiliations
Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand; Center of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand; Center of Innovative Immunodiagnostic Development, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand; Biomedical Technology Research Center, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency at the Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand
Type
Article
Source Title
Biosensors
ISSN
20796374
Year
2021
Volume
11
Issue
3
Open Access
All Open Access, Gold, Green
Publisher
MDPI
DOI
10.3390/bios11030076
Format
Abstract
Protein and DNA interactions are crucial for many cellular processes. Biolayer Interfer-ometry (BLI) is a label-free technology for determining kinetic biomolecular interactions with high accuracy results. In the present study, we determined the kinetic binding of a zinc finger scaffold, 2LTRZFP, which formerly constructed the interfering effect on HIV-1 integration process using BLI. The competitive Enzyme-linked immunosorbent assay (ELISA) was used to initially show the specific binding of 2LTRZFP. The percentages of inhibition were 62% and 22% in double-stranded 2LTR (ds2LTR) and irrelevant DNA (dsNeg), respectively. Consequently, the binding affinity of 2LTRZFP against ds2LTR target analyzed by BLI was 40 nM, which is stronger than the interaction of HIV-1 integrase (IN) enzyme to the 2LTR circle junction. Additionally, the 2LTRZFP did not interact with the genomic DNA extracted from SupT1 cell line. This result indicates that 2LTRZFP did not exhibit off-target effects against human genome. The knowledge obtained from this study supports the prospect of using 2LTRZFP in HIV-1 gene therapy. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
Keyword
2LTRZFP | Biolayer interferometry | HIV-1 | Zinc finger protein
Industrial Classification
Knowledge Taxonomy Level 1
Knowledge Taxonomy Level 2
Knowledge Taxonomy Level 3
Funding Sponsor
Office of the Higher Education Commission; National Science and Technology Development Agency; Higher Education Commission, Pakistan; National Research Council of Thailand
License
N/A
Rights
N/A
Publication Source
Scopus