2. Academic Validation
  3. Protoplast Isolation and Transfection in Wheat

Protoplast Isolation and Transfection in Wheat

  • Methods Mol Biol. 2022:2464:131-141. doi: 10.1007/978-1-0716-2164-6_10.
Guangbin Luo # 1,  Boshu Li # 2 3,  Caixia Gao 4 5
Affiliations

Affiliations

  • 1 NovoCrops Center, Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg C, Denmark.
  • 2 State Key Laboratory of Plant Cell and Chromosome Engineering, Center for Genome Editing, Institute of Genetics and Developmental Biology, Innovation Academy for Seed Design, Chinese Academy of Sciences, Beijing, China.
  • 3 College of Advanced Agricultural Sciences, University of Chinese Academy of Sciences, Beijing, China.
  • 4 State Key Laboratory of Plant Cell and Chromosome Engineering, Center for Genome Editing, Institute of Genetics and Developmental Biology, Innovation Academy for Seed Design, Chinese Academy of Sciences, Beijing, China. [email protected].
  • 5 College of Advanced Agricultural Sciences, University of Chinese Academy of Sciences, Beijing, China. [email protected].
  • # Contributed equally.
PMID: 35258830 DOI: 10.1007/978-1-0716-2164-6_10
Abstract

Wheat is one of the major staple crops around the world. A transient expression system is crucial for gene functional studies in wheat as stable transfection is still difficult in most cultivars. Protoplasts could serve as a versatile transient expression tool in wheat research. Here, we describe protocols for wheat protoplast isolation and transfection that are enabled by cellulase R-10 and macerozyme R-10 containing enzymatic solution and polyethylene glycol-mediated method, respectively. In addition, we show an example of efficiency evaluation of the emerging base editors in wheat protoplasts. These protocols are of wide use in both conventional gene functional analysis and reagent functionality evaluation of genome editing in wheat.

Keywords

Base editor; Base-editing efficiency; Genome editing; Polyethylene glycol (PEG); Triticum aestivum.

Figures
Products