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).

Generation of α-solanine-free hairy roots of potato by CRISPR/Cas9 mediated genome editing of the St16DOX gene

Typ / Jahr

Journal Article / 2018


Nakayasu, Masaru; Akiyama, Ryota; Lee, Hyoung Jae; Osakabe, Keishi; Osakabe, Yuriko; Watanabe, Bunta; Sugimoto, Yukihiro; Umemoto, Naoyuki; Saito, Kazuki; Muranaka, Toshiya; Mizutani, Masaharu


Potato (Solanum tuberosum) is a major food crop, while the most tissues of potato accumulates steroidal glycoalkaloids (SGAs) α-solanine and α-chaconine. Since SGAs confer a bitter taste on human and show the toxicity against various organisms, reducing the SGA content in the tubers is requisite for potato breeding. However, generation of SGA-free potato has not been achieved yet, although silencing of several SGA biosynthetic genes led a decrease in SGAs. Here, we show that the knockout of St16DOX encoding a steroid 16α-hydroxylase in SGA biosynthesis causes the complete abolition of the SGA accumulation in potato hairy roots. Nine candidate guide RNA (gRNA) target sequences were selected from St16DOX by in silico analysis, and the two or three gRNAs were introduced into a CRISPR/Cas9 vector designated as pMgP237-2A-GFP that can express multiplex gRNAs based on the pre-tRNA processing system. To establish rapid screening of the candidate gRNAs that can efficiently mutate the St16DOX gene, we used a potato hairy root culture system for the introduction of the pMgP237 vectors. Among the transgenic hairy roots, two independent lines showed no detectable SGAs but accumulated the glycosides of 22,26-dihydroxycholesterol, which is the substrate of St16DOX. Analysis of the two lines with sequencing exhibited the mutated sequences of St16DOX with no wild-type sequences. Thus, generation of SGAfree hairy roots of tetraploid potato was achieved by the combination of the hairy root culture and the pMgP237-2A-GFP vector. This experimental system is useful to evaluate the efficacy of candidate gRNA target sequences in the short-term.

16DOX; CRISPR/Cas9; Hairy roots; mutagenesis; Solanum tuberosum; Steroidal glycoalkaloids
Plant physiology and biochemistry : PPB
Periodical Number
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ID Corresponding Author
Plant Species GE Technique
Sequence Identifier
Type of Alteration
Progress in Research
Key Topic
781 Mizutani, Masaharu
Solanum tuberosum CRISPR/Cas9
Complete abolition of steroidal glycoalkaloids (SGAs) α-solanine and α-chaconine
Product quality