RNA-guided machineries perfectly satisfy the demand for rationally programmable tools that enable to precisely interfere with cellular gene expression. Since the last years it is becaming increasingly clear that there is an additional layer of information encoded into the RNA transcript referred to as epiranscriptomics. These included many chemical modification of transcriped message. To study and to harness such epitroanscirptomic events, we develop tools for the site-specific manipulation of epitranscriptomics marks, like inosine, for instance. With respect to the latter, we engineered artifical A-to-I editases based on the human protein ADAR. Since inosine is biochemically read as guanosine, A-to-I editing allows for reprogramming genetic information directly on the RNA-level. The natural editing process allows the cell to conditionally include point mutations into proteins by reprogramming single amino acid triplets on the mRNA. Thus protein isoforms with distinct properties are formed without the need for additional genomic copies. Furthermore, RNA processing signals including splice sites, miRNA binding sites, Start and Stop signals are targets of RNA editing.
Engineering deaminases (SNAP-ADARs)
Efficient and Precise Editing of Endogenous Transcripts with SNAP-tagged ADARs
P. Vogel, M. Moschref, Q. Li, T. Merkle, K. D. Selvasaravanan, J. B. Li, T. Stafforst*. Nature Methods 2018, 15, 535-38.
An RNA-Deaminase Conjugate Selectively Repairs Point Mutations
T. Stafforst*, M. F. Schneider, Angew. Chem. Int. Ed. 2012, 51, 11166-9.
Recruitment of endogenous ADARs
Precise RNA editing by recruiting endogenous ADARs with antisense oligonucleotides
T. Merkle, S. Merz, P. Reautschnig, A. Blaha, Q. Li, P. Vogel, J. Wettengel, J. B. Li, T. Stafforst*, Nature Biotech. 2019, in press
Harnessing human ADAR2 for RNA repair – Recoding a PINK1 mutation rescues mitophagy
J. Wettengel, P. Reautschnig, S. Geisler, P. J. Kahle, T. Stafforst*, Nucl. Acids Res. 2017, 45, 2797-2808.