Ribozyme-processed guide RNA enhances virus-mediated plant genome editing

Cited 2 time in scopus
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Title
Ribozyme-processed guide RNA enhances virus-mediated plant genome editing
Author(s)
Y Oh; H Kim; Hyo Jun Lee; S G Kim
Bibliographic Citation
Biotechnology Journal, vol. 17, no. 7, pp. 2100189-2100189
Publication Year
2022
Abstract
In virus-induced gene-editing system, subgenomic promoters have been used to express guide RNAs (gRNAs). However, the transcription initiation site of the subgenomic promoters remains elusive. Here, we examined the sequence of gRNAs expressed by subgenomic promoters and found the variable length of overhangs at 5'-end of gRNAs. The overhangs at 5'-end of gRNA decrease the cleavage activity of SpCas9. To overcome this problem, we inserted hammerhead ribozyme between the subgenomic promoter and gRNA and confirmed that gRNAs with a precise 5'-end increase the editing efficacy in wild tobacco. This system will be widely used for editing target genes in plants with high efficiency.
Keyword
CRISPR-Cas9Hammerhead ribozymePlant genome editingTobacco rattle virus vectorVirus-induced genome editing
ISSN
1860-6768
Publisher
Wiley
DOI
http://dx.doi.org/10.1002/biot.202100189
Type
Article
Appears in Collections:
Division of Research on National Challenges > Plant Systems Engineering Research > 1. Journal Articles
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