The ability to selectively interfere with post-translationally modified proteins would have many biological and therapeutic applications. However, post-translational modifications cannot be selectively targeted by nucleic-acid-based interference approaches. Here we describe post-translational intracellular silencing antibody technology (PISA), a method for selecting intrabodies against post-translationally modified proteins. We demonstrate our method by generating intrabodies against native acetylated proteins and showing functional interference in living cells.
Gene Expression Omnibus
NCBI Reference Sequence
The work was supported by Scuola Normale Superiore institutional funds and by a grant from the European Union Seventh Framework Program (grant no. 604102; Human Brain Project). We are grateful to M.-H. Kuo and D. Guo (Michigan State University, USA) for the H3 tethered catalysis bait, to A. Cereseto (University of Trento, Italy) for the HIV-1 integrase tethered catalysis bait. We acknowledge A. Allouch for helpful discussions about integrase protocols and V. Liverani for help in bait cloning and technical laboratory support. We thank T.H. Rabbitts, J. Zeng, and G. Meli for discussions on the yeast selection procedures. We also thank M. Maffei and R. Ciampi for collaborative logistic help. Many thanks to F. Cremisi and to A. Cellerino for helpful discussions.
Integrated supplementary information
Heat map of the significant differentially expressed genes between scFv112A (anti-acIN) and scFv58F (anti H3AcK9) samples.