TY - JOUR T1 - CRISPR-Cas12a-assisted PCR tagging of mammalian genes JF - bioRxiv DO - 10.1101/473876 SP - 473876 AU - Julia Fueller AU - Konrad Herbst AU - Matthias Meurer AU - Krisztina Gubicza AU - Bahtiyar Kurtulmus AU - Julia D. Knopf AU - Daniel Kirrmaier AU - Benjamin C. Buchmuller AU - Gislene Pereira AU - Marius K. Lemberg AU - Michael Knop Y1 - 2019/01/01 UR - http://biorxiv.org/content/early/2019/10/30/473876.abstract N2 - Here we describe a simple and time-efficient strategy for endogenous gene tagging in mammalian tissue culture cells. We take advantage of CRISPR-Cas12a-induced double strand breaks and their repair by homologous recombination using heterologous templates with short homology arms. Assembly of these templates and the Cas12a endonuclease is restricted to a single PCR step followed by DNA transfection, which avoids handling of RNAs, recombinant proteins or cloning of plasmids. The method is robust and works in all cell lines tested with tagging efficiency of up to 20% without selection, and up to 60% when selection markers are used. We show application in different cell lines and make a comprehensive set of C-terminal tagging constructs available to the research community. ER -