IFIB – Interfakultäres Institut für Biochemie

Research

Nucleic acids are fantastic molecules. Due to their unique base pairing properties, hybridization between nucleic acids is highly predictable and thus rationally programmable. We develop artifical, RNA-guided machines to create tools for the study and control of basic biochemical processes like RNA processing, translation, or epitranscriptomics. To achieve our goals, we combine chemistry with biotechnology and optimize their application inside cells and living organisms.

Site-directed RNA editing

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.

Key publications

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.

 

 

Photocontrol of biochemical processes

Light is an attractive external trigger for various processes as it enables a precise control in time, space and dosage. We are interested in controlling various biochemical processes by light. These include nucleic acid hybridization with psoralen crosslinks, ribonucleoprotein assembly via SNAP-tag technology, and photorelease of nitrogen oxide among others.

Key publication

Npom-protected NONOate enables light-triggered NO/cGMP signalling in primary vascular smooth muscle cells

A. S. Stroppel, M. Paolillo, T. Ziegler, R. Feil, T. Stafforst*. ChemBioChem 2018, 19, 1312-18.

 

Site-directed RNA editing in vivo can be triggered by the light-driven assembly of an artificial riboprotein

A. Hanswillemenke, T. Kuzdere, P. Vogel, G. Jekely, T. Stafforst*, J. Am. Chem. Soc. 2015, 137, 15 875-81.

 

Photoactivation of a Psoralen-blocked Luciferase Gene by Blue Light

T. Stafforst*, J. M. Stadler, Angew. Chem. Int. Ed. 2013, 52, 12448-51.