TY  - JOUR
AU  - Eggenschwiler, Reto
AU  - Gschwendtberger, Thomas
AU  - Felski, Christian
AU  - Jahn, Christopher
AU  - Langer, Florian
AU  - Sterneckert, Jared
AU  - Hermann, Andreas
AU  - Lühmann, Jonathan
AU  - Steinemann, Doris
AU  - Haase, Alexandra
AU  - Martin, Ulrich
AU  - Petri, Susanne
AU  - Cantz, Tobias
TI  - A selectable all-in-one CRISPR prime editing piggyBac transposon allows for highly efficient gene editing in human cell lines.
JO  - Scientific reports
VL  - 11
IS  - 1
SN  - 2045-2322
CY  - [London]
PB  - Macmillan Publishers Limited, part of Springer Nature
M1  - DZNE-2022-00385
SP  - 22154
PY  - 2021
AB  - 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
KW  - Amyotrophic Lateral Sclerosis: genetics
KW  - CRISPR-Cas Systems
KW  - Cell Line
KW  - Gene Editing: methods
KW  - HEK293 Cells
KW  - Humans
KW  - Induced Pluripotent Stem Cells: metabolism
KW  - Mutation
KW  - Superoxide Dismutase-1: genetics
KW  - Transfection: methods
KW  - Superoxide Dismutase-1 (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:34773059
C2  - pmc:PMC8589839
DO  - DOI:10.1038/s41598-021-01689-2
UR  - https://pub.dzne.de/record/163639
ER  -