TY  - INPR
A1  - Maghami, Mohammad Ghaem
A1  - Scheitl, Carolin P. M.
A1  - Höbartner, Claudia
T1  - Direct in vitro selection of trans-acting ribozymes for posttranscriptional, site-specific, and covalent fluorescent labeling of RNA
T2  - Journal of the American Chemical Society
N2  - General and efficient tools for site-specific fluorescent or bioorthogonal labeling of RNA are in high demand. Here, we report direct in vitro selection, characterization, and application of versatile trans-acting 2'-5' adenylyl transferase ribozymes for covalent and site-specific RNA labeling. The design of our partially structured RNA pool allowed for in vitro evolution of ribozymes that modify a predetermined nucleotide in cis (i.e. intramolecular reaction), and were then easily engineered for applications in trans (i.e. in an intermolecular setup). The resulting ribozymes are readily designed for specific target sites in small and large RNAs and accept a wide variety of N6-modified ATP analogues as small molecule substrates. The most efficient new ribozyme (FH14) shows excellent specificity towards its target sequence also in the context of total cellular RNA.
KW  - covalent and site-specific RNA labeling
KW  - trans-acting 2'-5' adenylyl transferase ribozymes
KW  - in vitro selection from a structured RNA library
KW  - Ribozyme-catalyzed RNA labeling
KW  - intermolecular applications of ribozymes
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-192333
N1  - This document is the unedited Author’s version of a Submitted Work that was subsequently accepted for publication in Journal of the American Chemical Society, copyright © American Chemical Society after peer review. To access the final edited and published work see https://doi.org/10.1021/jacs.9b10531.
ER  - 
TY  - JOUR
A1  - Maghami, Mohammad Ghaem
A1  - Dey, Surjendu
A1  - Lenz, Ann-Kathrin
A1  - Höbartner, Claudia
T1  - Repurpsing Antiviral Drugs for Orthogonal RNA-Catalyzed Labeling
JF  - Angewandte Chemie, International Edition
N2  - In vitro selected ribozymes are promising tools for site-specific labeling of RNA. Previously known nucleic acid catalysts attached fluorescently labeled adenosine or guanosine derivatives through 2’,5’-branched phosphodiester bonds to the RNA of interest. Herein, we report new ribozymes that use orthogonal substrates, derived from the antiviral drug tenofovir, and attach bioorthogonal functional groups, as well as affinity handles and fluorescent reporter units through a hydrolytically more stable phosphonate ester linkage. The tenofovir transferase ribozymes were identified by in vitro selection and are orthogonal to nucleotide transferase ribozymes. As genetically encodable functional RNAs, these ribozymes may be developed for potential cellular applications. The orthogonal ribozymes addressed desired target sites in large RNAs in vitro, as shown by fluorescent labeling of E. coli 16S and 23S RNAs in total cellular RNA.
KW  - Antiviral nucleoside analogues
KW  - in vitro selection
KW  - ribozymes
KW  - site-specific RNA labeling
KW  - tenofovir
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-205552
VL  - 59
ER  -