In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic SilencingAlessandro Migliara 1, Martino Alfredo Cappelluti 1, Francesca Giannese 2, Sara Valsoni 1, Alberto Coglot 1, Ivan Merelli 1,3, Davide Cittaro 2, Angelo Lombardo 1,4
1San Raffaele Telethon Institute for Gene Therapy (SR-Tiget), IRCCS San Raffaele Scientific Institute, 2Center for Omics Sciences, IRCCS San Raffaele Institute, 3Institute for Biomedical Technologies, National Research Council, 4Vita-Salute San Raffaele University
Here, we present a protocol for the in vitro selection of engineered transcriptional repressors (ETRs) with high, long-term, stable, on-target silencing efficiency and low genome-wide, off-target activity. This workflow allows for reducing an initial, complex repertoire of candidate ETRs to a short list, suitable for further evaluation in therapeutically relevant settings.
