Synthetic anti-RNA antibody derivatives for RNA visualization in mammalian cells

dc.contributor.authorBanna, Hasan Al
dc.contributor.authorBerg, Kimberley
dc.contributor.authorSadat, Tasnia
dc.contributor.authorDas, Naba Krishna
dc.contributor.authorPaudel, Roshan
dc.contributor.authorD'Souza, Victoria
dc.contributor.authorKoirala, Deepak
dc.date.accessioned2025-01-31T18:24:01Z
dc.date.available2025-01-31T18:24:01Z
dc.date.issued2024-12-31
dc.description.abstractAlthough antibody derivatives, such as Fabs and scFvs, have revolutionized the cellular imaging, quantification and tracking of proteins, analogous tools and strategies are unavailable for cellular RNA visualization. Here, we developed four synthetic anti-RNA scFv (sarabody) probes and their green fluorescent protein (GFP) fusions and demonstrated their potential to visualize RNA in live mammalian cells. We expressed these sarabodies and sarabody–GFP modules, purified them as soluble proteins, characterized their binding interactions with their corresponding epitopes and finally employed two of the four modules, sara1-GFP and sara1c-GFP, to visualize a target messenger RNA in live U2OS cells. Our current RNA imaging strategy is analogous to the existing MCP-MS₂ system for RNA visualization, but additionally, our approach provides robust flexibility for developing target RNA-specific imaging modules, as epitope-specific probes can be selected from a library generated by diversifying the sarabody complementarity determining regions. While we continue to optimize these probes, develop new probes for various target RNAs and incorporate other fluorescence proteins like mCherry and HaloTag, our groundwork results demonstrated that these first-of-a-kind immunofluorescent probes will have tremendous potential for tracking mature RNAs and may aid in visualizing and quantifying many cellular processes as well as examining the spatiotemporal dynamics of various RNAs.
dc.description.sponsorshipNational Institute of General Medical Sciences [1R35GM150869]; Division of Microbiology and Infectious Diseases [NIH NIAID1U54AI170660 and SUBK00019302]. Funding for open access charge: National Institute of General Medical Sciences [1R35GM150869].
dc.description.urihttps://academic.oup.com/nar/advance-article/doi/10.1093/nar/gkae1275/7935008#499083084
dc.format.extent14 pages
dc.genrejournal articles
dc.identifierdoi:10.13016/m2ordk-e0ha
dc.identifier.citationBanna, Hasan Al, Kimberley Berg, Tasnia Sadat, Naba Krishna Das, Roshan Paudel, Victoria D’Souza, and Deepak Koirala. "Synthetic Anti-RNA Antibody Derivatives for RNA Visualization in Mammalian Cells". Nucleic Acids Research (December 31, 2024): gkae1275. https://doi.org/10.1093/nar/gkae1275.
dc.identifier.urihttps://doi.org/10.1093/nar/gkae1275
dc.identifier.urihttp://hdl.handle.net/11603/37539
dc.language.isoen_US
dc.publisherOxford
dc.relation.isAvailableAtThe University of Maryland, Baltimore County (UMBC)
dc.relation.ispartofUMBC Staff Collection
dc.relation.ispartofUMBC Student Collection
dc.relation.ispartofUMBC Chemistry & Biochemistry Department
dc.rightsAttribution-NonCommercial 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by-nc/4.0/deed.en
dc.titleSynthetic anti-RNA antibody derivatives for RNA visualization in mammalian cells
dc.typeText
dcterms.creatorhttps://orcid.org/0000-0001-7475-6706
dcterms.creatorhttps://orcid.org/0000-0001-6424-3173
dcterms.creatorhttps://orcid.org/0000-0002-5308-2746

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