Multiplexed promoter and gene editing in wheat using a virus-based guide RNA delivery system.

Autor: Wang W; Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.; Wheat Genetic Resources Center, Kansas State University, Manhattan, KS, USA., Yu Z; Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.; Wheat Genetic Resources Center, Kansas State University, Manhattan, KS, USA., He F; Department of Plant Pathology, Kansas State University, Manhattan, KS, USA., Bai G; Hard Winter Wheat Genetics Research Unit, USDA-ARS, Manhattan, KS, USA., Trick HN; Department of Plant Pathology, Kansas State University, Manhattan, KS, USA., Akhunova A; Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.; Integrated Genomic Facility, Kansas State University, Manhattan, KS, USA., Akhunov E; Department of Plant Pathology, Kansas State University, Manhattan, KS, USA.; Wheat Genetic Resources Center, Kansas State University, Manhattan, KS, USA.
Jazyk: angličtina
Zdroj: Plant biotechnology journal [Plant Biotechnol J] 2022 Dec; Vol. 20 (12), pp. 2332-2341. Date of Electronic Publication: 2022 Sep 07.
DOI: 10.1111/pbi.13910
Abstrakt: The low efficiency of genetic transformation and gene editing across diverse cultivars hinder the broad application of CRISPR technology for crop improvement. The development of virus-based methods of CRISPR-Cas system delivery into the plant cells holds great promise to overcome these limitations. Here, we perform direct inoculation of wheat leaves with the barley stripe mosaic virus (BSMV) transcripts to deliver guide RNAs (sgRNA) into the Cas9-expressing wheat. We demonstrate that wheat inoculation with the pool of BSMV-sgRNAs could be used to generate heritable precise deletions in the promoter region of a transcription factor and to perform multiplexed editing of agronomic genes. We transfer the high-expressing locus of Cas9 into adapted spring and winter cultivars by marker-assisted introgression and use of the BSMV-sgRNAs to edit two agronomic genes. A strategy presented in our study could be applied to any adapted cultivar for creating new cis-regulatory diversity or large-scale editing of multiple genes in biological pathways or QTL regions, opening possibilities for the effective engineering of crop genomes, and accelerating gene discovery and trait improvement efforts.
(© 2022 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)
Databáze: MEDLINE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje