2LTRZFP Interacts Specifically to HIV-1 DNA without Off-Target Effects as Determined by Biolayer Interferometry
Protein and DNA interactions are crucial for many cellular processes. Biolayer Interferometry (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, whi...
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| Format: | Article |
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MDPI AG
2021-03-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/11/3/76 |
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| author | Koollawat Chupradit Weeraya Thongkum On-anong Juntit Kanokporn Sornsuwan Chatchai Tayapiwatana |
| author_facet | Koollawat Chupradit Weeraya Thongkum On-anong Juntit Kanokporn Sornsuwan Chatchai Tayapiwatana |
| author_sort | Koollawat Chupradit |
| collection | DOAJ |
| description | Protein and DNA interactions are crucial for many cellular processes. Biolayer Interferometry (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. |
| first_indexed | 2024-03-09T05:01:10Z |
| format | Article |
| id | doaj.art-7674e80fceff46b29eab41626100a861 |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-09T05:01:10Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-7674e80fceff46b29eab41626100a8612023-12-03T12:59:48ZengMDPI AGBiosensors2079-63742021-03-011137610.3390/bios110300762LTRZFP Interacts Specifically to HIV-1 DNA without Off-Target Effects as Determined by Biolayer InterferometryKoollawat Chupradit0Weeraya Thongkum1On-anong Juntit2Kanokporn Sornsuwan3Chatchai Tayapiwatana4Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, ThailandDivision of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, ThailandDivision of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, ThailandDivision of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, ThailandDivision of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, ThailandProtein and DNA interactions are crucial for many cellular processes. Biolayer Interferometry (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.https://www.mdpi.com/2079-6374/11/3/76HIV-1biolayer interferometryzinc finger protein2LTRZFP |
| spellingShingle | Koollawat Chupradit Weeraya Thongkum On-anong Juntit Kanokporn Sornsuwan Chatchai Tayapiwatana 2LTRZFP Interacts Specifically to HIV-1 DNA without Off-Target Effects as Determined by Biolayer Interferometry Biosensors HIV-1 biolayer interferometry zinc finger protein 2LTRZFP |
| title | 2LTRZFP Interacts Specifically to HIV-1 DNA without Off-Target Effects as Determined by Biolayer Interferometry |
| title_full | 2LTRZFP Interacts Specifically to HIV-1 DNA without Off-Target Effects as Determined by Biolayer Interferometry |
| title_fullStr | 2LTRZFP Interacts Specifically to HIV-1 DNA without Off-Target Effects as Determined by Biolayer Interferometry |
| title_full_unstemmed | 2LTRZFP Interacts Specifically to HIV-1 DNA without Off-Target Effects as Determined by Biolayer Interferometry |
| title_short | 2LTRZFP Interacts Specifically to HIV-1 DNA without Off-Target Effects as Determined by Biolayer Interferometry |
| title_sort | 2ltrzfp interacts specifically to hiv 1 dna without off target effects as determined by biolayer interferometry |
| topic | HIV-1 biolayer interferometry zinc finger protein 2LTRZFP |
| url | https://www.mdpi.com/2079-6374/11/3/76 |
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