Title

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

Authors

Authors

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

Comments

Authors: contact us about adding a copy of your work at STARS@ucf.edu

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

Share

COinS