Protein-PNA conjugates as conditional BRET platforms for life cell mRNA imaging and quantification
Final Report Abstract
During this project, novel approaches for the detection of low-copy mRNA transcripts in their native form were explored. Despite the relevance of mRNA isoforms in protein diversity and their inherent implication in diseases (above all in neurological ones), real-time visualization of mRNA isoforms in living cells has been hampered due to the lack of sensitivity of existing methodologies. Taking the well-characterized alternative splicing of the survival motor neuron (SMN2) gene as a starting point, different strategies based on bioluminescence were tested for this purpose. In particular, peptide-nucleic acids (PNAs) in combination with the NanoLuc Luciferase demonstrated to specifically recognise and visualize mRNA transcripts in living cells, surpassing the detection of the most sensitive reported aptamers. Thus, native bioluminescent mRNA was visualized in living cells without needing washing, or transfection steps. Notably, owing to the PNA programmability, this approach can be tailored to detect any desirable target. Consequently, this mRNA sensor represents a significant advancement, and further applications are expected.
Publications
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Bistable Photoswitch Allows in Vivo Control of Hematopoiesis. ACS Central Science, 8(1), 57-66.
Albert, Lea; Nagpal, Jatin; Steinchen, Wieland; Zhang, Lei; Werel, Laura; Djokovic, Nemanja; Ruzic, Dusan; Hoffarth, Malte; Xu, Jing; Kaspareit, Johanna; Abendroth, Frank; Royant, Antoine; Bange, Gert; Nikolic, Katarina; Ryu, Soojin; Dou, Yali; Essen, Lars-Oliver & Vázquez, Olalla
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A Chemical Biology Perspective to Therapeutic Regulation of RNA Splicing in Spinal Muscular Atrophy (SMA). ACS Chemical Biology, 17(6), 1293-1307.
Zhang, Lei; Abendroth, Frank & Vázquez, Olalla
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Synthesis of the l-and d-SH2 domain of the leukaemia oncogene Bcr-Abl. RSC Chemical Biology, 3(8), 1008-1012.
Schmidt, Nina; Abendroth, Frank; Vázquez, Olalla & Hantschel, Oliver
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Cell-free biosynthesis combined with deep learning accelerates de novo-development of antimicrobial peptides. Nature Communications, 14(1).
Pandi, Amir; Adam, David; Zare, Amir; Trinh, Van Tuan; Schaefer, Stefan L.; Burt, Marie; Klabunde, Björn; Bobkova, Elizaveta; Kushwaha, Manish; Foroughijabbari, Yeganeh; Braun, Peter; Spahn, Christoph; Preußer, Christian; Pogge von Strandmann, Elke; Bode, Helge B.; von Buttlar, Heiner; Bertrams, Wilhelm; Jung, Anna Lena; Abendroth, Frank ... & Erb, Tobias J.
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Peptide-mediated inhibition of the transcriptional regulator Elongin BC induces apoptosis in cancer cells. Cell Chemical Biology, 30(7), 766-779.e11.
Fischer, Sabrina; Trinh, Van Tuan; Simon, Clara; Weber, Lisa M.; Forné, Ignasi; Nist, Andrea; Bange, Gert; Abendroth, Frank; Stiewe, Thorsten; Steinchen, Wieland; Liefke, Robert & Vázquez, Olalla
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Reversible Control of RNA Splicing by Photoswitchable Small Molecules. Journal of the American Chemical Society, 145(23), 12783-12792.
Zhang, Lei; Xie, Xiulan; Djokovic, Nemanja; Nikolic, Katarina; Kosenkov, Dmitri; Abendroth, Frank & Vázquez, Olalla
