Cell-type and activation-specific labelling of leukocytes through bioorthogonal tagging in vivo

Principal investigators: Andrea Rentmeister, Henning Mootz
Project number: CRC 1450 A01
Project term: 01/2021–12/2024

Graphical abstract
© CRC inSight

Imaging of different leukocyte populations in vivo is key to understand their dynamics and role in inflammation. To overcome current limitations, we will develop new methods to label immune cells (1) within a longer time window, i.e., from initiation of inflammation to resolution. We will establish a flexible and modular platform applicable to different imaging modalities, such as fluorescence, scintigraphic or photoacoustic imaging. ER-HoxB8 cells (2) will be genetically tagged ex vivo with self-labelling enzymes or epitopes for antibody fragments (3). After differentiation and re-injection, in vivo labelling (4) will be achieved by administering tag-specific, functionalized labels to selectively visualize the respective cell population. Novel approaches will be initially evaluated in a cutaneous granuloma mouse model and then expanded to other disease models in CRC projects.


Principal investigators

Project members


The names of the principal investigators in our network have been bolded. Publications released prior to 2021, when funding for our network commenced, represent previous project-related work.


Depke DA, Konken CP, Rosner L, Hermann S, Schafers M, Rentmeister A. A novel (18)F-labeled clickable substrate for targeted imaging of SNAP-tag expressing cells by PET in vivo. Chem Commun (Camb) 2021;57: 9850-9853. Abstract


Jedlitzke B, Yilmaz Z, Dorner W, Mootz HD. Photobodies: Light-Activatable Single-Domain Antibody Fragments. Angew Chem Int Ed Engl 2020;59: 1506-1510. Abstract
Westerich KJ, Chandrasekaran KS, Gross-Thebing T, Kueck N, Raz E, Rentmeister A. Bioorthogonal mRNA labeling at the poly(A) tail for imaging localization and dynamics in live zebrafish embryos. Chem. Sci. 2020;11: 3089-3095. Abstract


Anhauser L, Huwel S, Zobel T, Rentmeister A. Multiple covalent fluorescence labeling of eukaryotic mRNA at the poly(A) tail enhances translation and can be performed in living cells. Nucleic Acids Res 2019;47: e42. Abstract
Bhagawati M, Terhorst TME, Fusser F, Hoffmann S, Pasch T, Pietrokovski S, Mootz HD. A mesophilic cysteine-less split intein for protein trans-splicing applications under oxidizing conditions. Proc Natl Acad Sci U S A 2019;116: 22164-22172. Abstract


Pirzer T, Becher K-S, Rieker M, Meckel T, Mootz HD, Kolmar H. Generation of Potent Anti-HER1/2 Immunotoxins by Protein Ligation Using Split Inteins. ACS Chem Biol 2018;13: 2058-2066. Abstract


Bachmann A-L, Mootz HD. N-terminal chemical protein labeling using the naturally split GOS-TerL intein. J Pept Sci 2017;23: 624-630. Abstract
Muttach F, Muthmann N, Reichert D, Anhäuser L, Rentmeister A. A benzylic linker promotes methyltransferase catalyzed norbornene transfer for rapid bioorthogonal tetrazine ligation. Chem. Sci. 2017;8: 7947-7953. Abstract


Holstein JM, Anhauser L, Rentmeister A. Modifying the 5'-Cap for Click Reactions of Eukaryotic mRNA and To Tune Translation Efficiency in Living Cells. Angew Chem Int Ed Engl 2016;55: 10899-10903. Abstract


Holstein JM, Stummer D, Rentmeister A. Enzymatic modification of 5′-capped RNA with a 4-vinylbenzyl group provides a platform for photoclick and inverse electron-demand Diels–Alder reaction. Chem. Sci. 2015;6: 1362-1369. Abstract


Thiel IV, Volkmann G, Pietrokovski S, Mootz HD. An atypical naturally split intein engineered for highly efficient protein labeling. Angew Chem Int Ed Engl 2014;53: 1306-1310. Abstract