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Research article - Peer-reviewed, 2019

High efficacy full allelic CRISPR/Cas9 gene editing in tetraploid potato

Johansen, Ida Elisabeth; Liu, Ying; Jorgensen, Bodil; Bennett, Eric Paul; Andreasson, Erik; Nielsen, Kare L.; Blennow, Andreas; Petersen, Bent Larsen;

Abstract

CRISPR/Cas9 editing efficacies in tetraploid potato were highly improved through the use of endogenous potato U6 promoters. Highly increased editing efficiencies in the Granular Bound Starch Synthase gene at the protoplast level were obtained by replacement of the Arabidopsis U6 promotor, driving expression of the CRISPR component, with endogenous potato U6 promotors. This translated at the ex-plant level into 35% full allelic gene editing. Indel Detection Amplicon Analysis was established as an efficient tool for fast assessment of gene editing in complex genomes, such as potato. Together, this warrants significant reduction of laborious cell culturing, ex-plant regeneration and screening procedures of plants with high complexity genomes.

Keywords

Nuclear RNA gene; guide RNA; genome; expression; sequence; Arabidopsis; prediction; resistance; mutation; cloning

Published in

Scientific Reports

2019, volume: 9, article number: 17715

Authors' information

Johansen, Ida Elisabeth
University of Copenhagen
Liu, Ying
University of Copenhagen
Jørgensen, Bodil
University of Copenhagen
Bennett, Eric Paul
University of Copenhagen
Swedish University of Agricultural Sciences, Department of Plant Protection Biology
Nielsen, Kåre L.
Aalborg University
Blennow, Andreas
University of Copenhagen
Larsen Petersen, Bent
Royal Veterinary and Agricultural University

UKÄ Subject classification

Plant Biotechnology

Publication Identifiers

DOI: https://doi.org/10.1038/s41598-019-54126-w

URI (permanent link to this page)

https://res.slu.se/id/publ/102773