RT Journal Article SR Electronic T1 CRISPR-Cas12a-assisted PCR tagging of mammalian genes JF bioRxiv FD Cold Spring Harbor Laboratory SP 473876 DO 10.1101/473876 A1 Julia Fueller A1 Konrad Herbst A1 Matthias Meurer A1 Krisztina Gubicza A1 Bahtiyar Kurtulmus A1 Julia D. Knopf A1 Daniel Kirrmaier A1 Benjamin C. Buchmuller A1 Gislene Pereira A1 Marius K. Lemberg A1 Michael Knop YR 2019 UL http://biorxiv.org/content/early/2019/10/30/473876.abstract AB 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.