Specific RNA-Binding Antibodies with a Four-Amino-Acid Code

    Authors

    E. M. Sherman; S. Holmes;J. D. Ye

    Comments

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    Abbreviated Journal Title

    J. Mol. Biol.

    Keywords

    NONCODING RNAS; SYNTHETIC ANTIBODIES; ULTRACONSERVED REGIONS; GLYCINE; RIBOSWITCH; STRUCTURAL BASIS; LIGAND-BINDING; RECOGNITION; CRYSTALLIZATION; EXPRESSION; LIBRARIES; Biochemistry & Molecular Biology

    Abstract

    Numerous large non-coding RNAs are rapidly being discovered, and many of them have been shown to play vital roles in gene expression, gene regulation, and human diseases. Given their often structured nature, specific recognition with an antibody fragment becomes feasible and may help define the structure and function of these non-coding RNAs. As demonstrated for protein antigens, specific antibodies may aid in RNA crystal structure elucidation or the development of diagnostic tools and therapeutic drugs targeting disease-causing RNAs. Recent success and limitation of RNA antibody development has made it imperative to generate an effective antibody library specifically targeting RNA molecules. Adopting the reduced chemical diversity design and further restricting the interface diversity to tyrosines, serines, glycines, and arginines only, we have constructed a RNA-targeting Fab library. Phage display selection and downstream characterization showed that this library yielded high-affinity Fabs for all three RNA targets tested. Using a quantitative specificity assay, we found that these Fabs are highly specific, possibly due to the alternate codon design we used to avoid consecutive arginines in the Fab interface. In addition, the effectiveness of the minimal Fab library may challenge our view of the protein-RNA binding interface and provide a unique solution for future design of RNA-binding proteins. (C) 2014 Elsevier Ltd. All rights reserved.

    Journal Title

    Journal of Molecular Biology

    Volume

    426

    Issue/Number

    10

    Publication Date

    1-1-2014

    Document Type

    Article

    Language

    English

    First Page

    2145

    Last Page

    2157

    WOS Identifier

    WOS:000336349000010

    ISSN

    0022-2836

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