Repositorium

What is a repositorium?

The repositorium is a searchable database that provides data on relevant articles from journals, company web pages and web pages of governmental agencies about studies/applications of genome-editing in model plants and agricultural crops in the period January 1996 to May 2018. Search options are article type, technique, plant, traits or free text. The repositorium is based on the systematic map of Dominik Modrzejewski et al., published in the journal environmental evidence. (Download article PDF).

Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single-cell mutation detection to mutant plant regeneration

449

Typ / Jahr

Journal Article / 2018

Autoren

Lin, Choun-Sea; Hsu, Chen-Tran; Yang, Ling-Hung; Lee, Lan-Ying; Fu, Jin-Yuan; Cheng, Qiao-Wei; Wu, Fu-Hui; Hsiao, Han C-W; Zhang, Yesheng; Zhang, Ru; Chang, Wan-Jung; Yu, Chen-Ting; Wang, Wen; Liao, Li-Jen; Gelvin, Stanton B.; Shih, Ming-Che

Abstract

Plant protoplasts are useful for assessing the efficiency of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) mutagenesis. We improved the process of protoplast isolation and transfection of several plant species. We also developed a method to isolate and regenerate single mutagenized Nicotianna tabacum protoplasts into mature plants. Following transfection of protoplasts with constructs encoding Cas9 and sgRNAs, target gene DNA could be amplified for further analysis to determine mutagenesis efficiency. We investigated N. tabacum protoplasts and derived regenerated plants for targeted mutagenesis of the phytoene desaturase (NtPDS) gene. Genotyping of albino regenerants indicated that all four NtPDS alleles were mutated in amphidiploid tobacco, and no Cas9 DNA could be detected in most regenerated plants.

Keywords

CRISPR/Cas9; protoplast isolation; protoplast regeneration; single-cell analysis.

Periodical

Plant biotechnology journal

Periodical Number

Page range

1295–1310

Volume

DOI

10.1111/pbi.12870

View Publication

Techniques

ID	Corresponding Author Country	Plant Species	GE Technique Sequence Identifier	Trait Type of Alteration	Progress in Research Key Topic
1018	Gelvin, Stanton B; Shih, Ming-Che Taiwan; USA	Arabidopsis thaliana	CRISPR/Cas9 PDS	Albino phenotype SDN1	Basic research Basic research
1019	Gelvin, Stanton B; Shih, Ming-Che Taiwan; USA	Bambus oldhamii	CRISPR/Cas9 PDS	Albino phenotype SDN1	Basic research Basic research
1020	Gelvin, Stanton B; Shih, Ming-Che Taiwan; USA	Setaria italica	CRISPR/Cas9 PDS	Albino phenotype SDN1	Basic research Basic research
1021	Gelvin, Stanton B; Shih, Ming-Che Taiwan; USA	Brassica oleracea	CRISPR/Cas9 GA4a	No information SDN1	Basic research Basic research
1022	Gelvin, Stanton B; Shih, Ming-Che Taiwan; USA	Zea mays	CRISPR/Cas9 IPK	No information SDN1	Basic research Basic research
1023	Gelvin, Stanton B; Shih, Ming-Che Taiwan; USA	Brassica napus	CRISPR/Cas9 GA4a	No information SDN1	Basic research Basic research
1024	Gelvin, Stanton B; Shih, Ming-Che Taiwan; USA	Oryza sativa	CRISPR/Cas9 PDS	Albino phenotype SDN1	Basic research Basic research
1025	Gelvin, Stanton B; Shih, Ming-Che Taiwan; USA	Nicotiana tabacum	CRISPR/Cas9 PDS	Albino phenotype SDN1	Basic research Basic research
1026	Gelvin, Stanton B; Shih, Ming-Che Taiwan; USA	Solanum lycopersicum	CRISPR/Cas9 PDS	Albino phenotype SDN1	Basic research Basic research