ABSTRACT
OBJECTIVE: To obtain transgene-free progeny by constructing a DNA editing system with a fluorescent screening marker gene and two pairs of single-guide RNAs to simultaneously recognize two different sites in the target gene encoding Arabidopsis microRNA(miR)160A RESULTS: The T1 seeds with red fluorescence were easily identified and were selected to verify that the proportion of miR160A knockout mutants reached approximately 50%. Seeds with no fluorescence in the T2 generation were selected and screened for homozygous mutants. In the T2 generation plants, the Cas9 fragment was not detected by polymerase chain reaction. The traits of the homozygous mutants were stably inherited by the T2 population. CONCLUSIONS: A highly efficient DNA-editing construct was successfully developed and can be used as a plant genome site-specific editing tool that may be useful for improving plant genetic resources.
Subject(s)
Arabidopsis/growth & development , Luminescent Proteins/metabolism , MicroRNAs/genetics , Arabidopsis/genetics , Gene Editing , Genetic Markers , Luminescent Proteins/genetics , MicroRNAs/metabolism , Mutation , RNA, Guide, Kinetoplastida , Transgenes , Red Fluorescent ProteinABSTRACT
The article An efficient transgene-free DNA-editing system for Arabidopsis using a fluorescent marker, written by Lejun Ouyang . Mingsai Ma and Limei Li, was originally published electronically on the publisher's internet portal (currently SpringerLink) on 03 December 2019 with open access. With the author(s)' decision to step back from Open Choice, the copyright of the article changed on 18 December 2019 to © Springer Nature B.V. 2019 and the article is forthwith distributed under the terms of copyright. The original article has been corrected.