Use of CRISPR/Cas9 for Targeted Mutagenesis in Sorghum.

Autor: Char SN; Division of Plant Sciences, University of Missouri, Columbia, Missouri.; Christopher S. Bond Life Sciences Center, University of Missouri, Columbia, Missouri., Lee H; Division of Plant Sciences, University of Missouri, Columbia, Missouri.; Plant Transformation Core Facility, University of Missouri, Columbia, Missouri., Yang B; Division of Plant Sciences, University of Missouri, Columbia, Missouri.; Christopher S. Bond Life Sciences Center, University of Missouri, Columbia, Missouri.; Donald Danforth Plant Science Center, St. Louis, Missouri.
Jazyk: angličtina
Zdroj: Current protocols in plant biology [Curr Protoc Plant Biol] 2020 Jun; Vol. 5 (2), pp. e20112.
DOI: 10.1002/cppb.20112
Abstrakt: Sorghum (Sorghum bicolor) fulfills the demand for bioenergy resources and also provides substantial diet calories to the world's population. Therefore, many biological studies use sorghum as a research model for improvement of the domesticated food and bioenergy crops. Furthermore, leveraging genome editing systems in a plethora of grass plant species has been extensively studied with no exception in sorghum. However, a protocol that details the genome editing strategies using CRISPR/Cas9 and that combines an efficient tissue culture and transformation platform in sorghum based on Agrobacterium-mediated DNA transfer has yet to be reported. This protocol outlines the steps and workflow from design of sorghum CRISPR target sites using BTx623 as a reference genome, construction of sorghum CRISPR/Cas9 plasmids, tissue culture, to Agrobacterium-mediated transformation followed by genotyping of CRISPR/Cas9 induced mutants. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Construction of CRISPR/Cas9 expression vector to analysis of CRISPR-edited plants Basic Protocol 2: Stable transformation of sorghum Support Protocol: Management of sorghum plants in a greenhouse.
(© 2020 Wiley Periodicals LLC.)
Databáze: MEDLINE
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