Site-Selective RNA Splicing Nanozyme: DNAzyme and RtcB Conjugates on a Gold Nanoparticle

• Type of Material: Article
• Authors: Jessica R. Petree, Emory University; Kevin Yehl, Emory University; Kornelia Galior, Emory University; Roxanne Glazier, Georgia Institute of Technology; Brendan Deal, Emory University; Khalid Salaita, Emory University
• Language: English
• Date: 2018-01-01
• Publisher: American Chemical Society
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2017 American Chemical Society.
• Final Published Version (URL): http://dx.doi.org/10.1021/acschembio.7b00437
• Title of Journal or Parent Work: ACS Chemical Biology
• ISSN: 1554-8929
• Volume: 13
• Issue: 1
• Start Page: 215
• End Page: 224
• Grant/Funding Information: J.P. would like to thank the ARCS Foundation and S. and F. Burke for their generous support.; R.G. would like to thank NIH (NIGMS GM124472) and the NSF Fellowship for financial support.; K.S. would like to thank the NIH (R01-GM097399) and the NSF CAREER Award (1350829) for financial support.; NSF CAREER Award and NIH
• Keywords: Life Sciences & Biomedicine; GENE-THERAPY; DNA; Biochemistry & Molecular Biology; Science & Technology; CELLS; IN-VIVO; 3'-PHOSPHATE; ARTIFICIAL ENZYMES; SEQUENTIAL 2',3'-CYCLIC PHOSPHODIESTERASE; RIBOZYME; LIGASE; LIGATION
• Research Categories: Chemistry, Biochemistry; Biology, Genetics; Biology, Molecular

Abstract

Modifying RNA through either splicing or editing is a fundamental biological process for creating protein diversity from the same genetic code. Developing novel chemical biology tools for RNA editing has potential to transiently edit genes and to provide a better understanding of RNA biochemistry. Current techniques used to modify RNA include the use of ribozymes, adenosine deaminase, and tRNA endonucleases. Herein, we report a nanozyme that is capable of splicing virtually any RNA stem-loop. This nanozyme is comprised of a gold nanoparticle functionalized with three enzymes: two catalytic DNA strands with ribonuclease function and an RNA ligase. The nanozyme cleaves and then ligates RNA targets, performing a splicing reaction that is akin to the function of the spliceosome. Our results show that the three-enzyme reaction can remove a 19 nt segment from a 67 nt RNA loop with up to 66% efficiency. The complete nanozyme can perform the same splice reaction at 10% efficiency. These splicing nanozymes represent a new promising approach for gene manipulation that has potential for applications in living cells.