Development of a single-domain antibody to target a G-quadruplex located on the hepatitis B virus covalently closed circular DNA genome

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dc.contributor.authorFigueroa, Gerardo B.
dc.contributor.authorD'souza, Simmone
dc.contributor.authorPereira, Higor S.
dc.contributor.authorVasudeva, Gunjan
dc.contributor.authorFigueroa, Sara B.
dc.contributor.authorRobinson, Zachary E.
dc.contributor.authorBadmalia, Maulik D.
dc.contributor.authorMeier-Stephenson, Vanessa
dc.contributor.authorCorcoran, Jennifer A.
dc.contributor.authorvan Marle, Guido
dc.contributor.authorNi, Yi
dc.contributor.authorUrban, Stephan
dc.contributor.authorCoffin, Carla S.
dc.contributor.authorPatel, Trushar R.
dc.date.accessioned2024-07-10T18:18:09Z
dc.date.available2024-07-10T18:18:09Z
dc.date.issued2024
dc.descriptionOpen access article. Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license (CC BY-NC-ND 4.0) applies
dc.description.abstractTo achieve a virological cure for hepatitis B virus (HBV), innovative strategies are required to target the covalently closed circular DNA (cccDNA) genome. Guanine-quadruplexes (G4s) are a secondary structure that can be adopted by DNA and play a significant role in regulating viral replication, transcription, and translation. Antibody-based probes and small molecules have been developed to study the role of G4s in the context of the human genome, but none have been specifically made to target G4s in viral infection. Herein, we describe the development of a humanized single-domain antibody (S10) that can target a G4 located in the PreCore (PreC) promoter of the HBV cccDNA genome. MicroScale Thermophoresis demonstrated that S10 has a strong nanomolar affinity to the PreC G4 in its quadruplex form and a structural electron density envelope of the complex was determined using Small-Angle X-ray Scattering. Lentiviral transduction of S10 into HepG2-NTCP cells shows nuclear localization, and chromatin immunoprecipitation coupled with next-generation sequencing demonstrated that S10 can bind to the HBV PreC G4 present on the cccDNA. This research validates the existence of a G4 in HBV cccDNA and demonstrates that this DNA secondary structure can be targeted with high structural and sequence specificity using S10.
dc.identifier.citationFigueroa, G. B., D'souza, S., Pereira, H. S., Vasudeva, G., Figueroa, S. B., Robinson, Z. E., Badmalia, M. D., Meier-Stephenson, V., Corcoran, J. A., van Marle, G., Ni, Yi, Urban, S., Coffin, C. S., & Patel, T. R. (2024). Development of a single-domain antibody to target a G-quadruplex located on the hepatitis B virus covalently closed circular DNA genome. Journal of Medical Virology, 96(6), Article e29692. https://doi.org/10.1002/jmv.29692
dc.identifier.urihttps://hdl.handle.net/10133/6810
dc.language.isoen
dc.publisherAmerican Society for Microbiology
dc.publisher.departmentDepartment of Chemistry and Biochemistry
dc.publisher.facultyArts and Science
dc.publisher.institutionUniversity of Lethbridge
dc.publisher.institutionUniversity of Calgary
dc.publisher.institutionUniversity of Alberta
dc.publisher.institutionUniversity Hospital Heidelberg
dc.publisher.institutionHeidelberg University
dc.subjectcccDNA chromatin immunoprecipitation
dc.subjectCovalently closed circular DNA
dc.subjectG-quadruplex
dc.subjectGuanine-quadruplex
dc.subjectHepatitis B virus
dc.subjectSingle-domain antibody
dc.subject.lcshCircular DNA
dc.subject.lcshQuadruplex nucleic acids
dc.titleDevelopment of a single-domain antibody to target a G-quadruplex located on the hepatitis B virus covalently closed circular DNA genome
dc.typeArticle
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