Protoplast isolation and regeneration from Gracilaria changii (Gracilariales, Rhodophyta)

2009 ◽  
pp. 191-201
Author(s):  
Hui-Yin Yeong ◽  
Norzulaani Khalid ◽  
Siew-Moi Phang
Keyword(s):  
Protoplast Isolation ◽  
Gracilaria Changii

Protoplast isolation and regeneration from Gracilaria changii (Gracilariales, Rhodophyta)

2007 ◽  
Vol 20 (5) ◽  
pp. 641-651 ◽  
Author(s):  
Hui-Yin Yeong ◽  
Norzulaani Khalid ◽  
Siew-Moi Phang
Keyword(s):  
Protoplast Isolation ◽  
Gracilaria Changii

Protoplast Isolation and Fusion Induced by PEG with Leaves and Roots of Tea Plant (Camellia sinensis L. O. Kuntze)

2018 ◽  
Vol 44 (3) ◽  
pp. 463 ◽  
Author(s):  
Zhang PENG ◽  
Hua-Rong TONG ◽  
Guo-Lu LIANG ◽  
Yi-Qi SHI ◽  
Lian-Yu YUAN
Keyword(s):  
Camellia Sinensis ◽  
Protoplast Isolation ◽  
Tea Plant ◽  
Leaves And Roots

Callus induction, suspension culture and protoplast isolation in Camellia oleifera

2021 ◽  
Vol 286 ◽  
pp. 110193
Author(s):  
Su-Fang Li ◽  
Tian-Wen Ye ◽  
Xin Xu ◽  
De-Yi Yuan ◽  
Shi-Xin Xiao
Keyword(s):  
Suspension Culture ◽  
Callus Induction ◽  
Protoplast Isolation ◽  
Camellia Oleifera

scholarly journals Optimization of Protoplast Isolation and Transformation for a Pilot Study of Genome Editing in Peanut by Targeting the Allergen Gene Ara h 2

2022 ◽  
Vol 23 (2) ◽  
pp. 837
Author(s):  
Sudip Biswas ◽  
Nancy J. Wahl ◽  
Michael J. Thomson ◽  
John M. Cason ◽  
Bill F. McCutchen ◽  
...  
Keyword(s):  
Genome Editing ◽  
New Technologies ◽  
Fluorescent Protein ◽  
Peanut Butter ◽  
Processing System ◽  
Protoplast Isolation ◽  
Sequencing Analysis ◽  
Ara H 2

The cultivated peanut (Arachis hypogaea L.) is a legume consumed worldwide in the form of oil, nuts, peanut butter, and candy. Improving peanut production and nutrition will require new technologies to enable novel trait development. Clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9 (CRISPR–Cas9) is a powerful and versatile genome-editing tool for introducing genetic changes for studying gene expression and improving crops, including peanuts. An efficient in vivo transient CRISPR–Cas9- editing system using protoplasts as a testbed could be a versatile platform to optimize this technology. In this study, multiplex CRISPR–Cas9 genome editing was performed in peanut protoplasts to disrupt a major allergen gene with the help of an endogenous tRNA-processing system. In this process, we successfully optimized protoplast isolation and transformation with green fluorescent protein (GFP) plasmid, designed two sgRNAs for an allergen gene, Ara h 2, and tested their efficiency by in vitro digestion with Cas9. Finally, through deep-sequencing analysis, several edits were identified in our target gene after PEG-mediated transformation in protoplasts with a Cas9 and sgRNA-containing vector. These findings demonstrated that a polyethylene glycol (PEG)-mediated protoplast transformation system can serve as a rapid and effective tool for transient expression assays and sgRNA validation in peanut.

Sign in / Sign up

Export Citation Format

Share Document