%0 Journal Article
%A Eggenschwiler, Reto
%A Gschwendtberger, Thomas
%A Felski, Christian
%A Jahn, Christopher
%A Langer, Florian
%A Sterneckert, Jared
%A Hermann, Andreas
%A Lühmann, Jonathan
%A Steinemann, Doris
%A Haase, Alexandra
%A Martin, Ulrich
%A Petri, Susanne
%A Cantz, Tobias
%T A selectable all-in-one CRISPR prime editing piggyBac transposon allows for highly efficient gene editing in human cell lines.
%J Scientific reports
%V 11
%N 1
%@ 2045-2322
%C [London]
%I Macmillan Publishers Limited, part of Springer Nature
%M DZNE-2022-00385
%P 22154
%D 2021
%X CRISPR prime-editors are emergent tools for genome editing and offer a versatile alternative approach to HDR-based genome engineering or DNA base-editors. However, sufficient prime-editor expression levels and availability of optimized transfection protocols may affect editing efficiencies, especially in hard-to-transfect cells like hiPSC. Here, we show that piggyBac prime-editing (PB-PE) allows for sustained expression of prime-editors. We demonstrate proof-of-concept for PB-PE in a newly designed lentiviral traffic light reporter, which allows for estimation of gene correction and defective editing resulting in indels, based on expression of two different fluorophores. PB-PE can prime-edit more than 50
%K Amyotrophic Lateral Sclerosis: genetics
%K CRISPR-Cas Systems
%K Cell Line
%K Gene Editing: methods
%K HEK293 Cells
%K Humans
%K Induced Pluripotent Stem Cells: metabolism
%K Mutation
%K Superoxide Dismutase-1: genetics
%K Transfection: methods
%K Superoxide Dismutase-1 (NLM Chemicals)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:34773059
%2 pmc:PMC8589839
%R 10.1038/s41598-021-01689-2
%U https://pub.dzne.de/record/163639