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dc.contributor.authorXu, Ziyan
dc.contributor.authorKuang, Yongjie
dc.contributor.authorRen, Bin
dc.contributor.authorYan, Daqi
dc.contributor.authorYan, Fang
dc.contributor.authorSpetz, Carl
dc.contributor.authorSun, Wenxian
dc.contributor.authorWang, Guirong
dc.contributor.authorZhou, Xueping
dc.contributor.authorZhou, Huanbin
dc.date.accessioned2021-11-19T09:04:48Z
dc.date.available2021-11-19T09:04:48Z
dc.date.created2021-03-29T21:33:52Z
dc.date.issued2021-01-04
dc.identifier.citationGenome Biology. 2021, 22 .en_US
dc.identifier.issn1465-6906
dc.identifier.urihttps://hdl.handle.net/11250/2830416
dc.description.abstractBackground Plant genome engineering mediated by various CRISPR-based tools requires specific protospacer adjacent motifs (PAMs), such as the well-performed NGG, NG, and NNG, to initiate target recognition, which notably restricts the editable range of the plant genome. Results In this study, we thoroughly investigate the nuclease activity and the PAM preference of two structurally engineered SpCas9 variants, SpG and SpRY, in transgenic rice. Our study shows that SpG nuclease favors NGD PAMs, albeit less efficiently than the previously described SpCas9-NG, and that SpRY nuclease achieves efficient editing across a wide range of genomic loci, exhibiting a preference of NGD as well as NAN PAMs. Furthermore, SpRY-fused cytidine deaminase hAID*Δ and adenosine deaminase TadA8e are generated, respectively. These constructs efficiently induce C-to-T and A-to-G conversions in the target genes toward various non-canonical PAMs, including non-G PAMs. Remarkably, high-frequency self-editing events (indels and DNA fragments deletion) in the integrated T-DNA fragments as a result of the nuclease activity of SpRY are observed, whereas the self-editing of SpRY nickase-mediated base editor is quite low in transgenic rice lines. Conclusions The broad PAM compatibility of SpRY greatly expands the targeting scope of CRISPR-based tools in plant genome engineering.en_US
dc.language.isoengen_US
dc.publisherBMCen_US
dc.rightsNavngivelse 4.0 Internasjonal*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/deed.no*
dc.titleSpRY greatly expands the genome editing scope in rice with highly flexible PAM recognitionen_US
dc.typePeer revieweden_US
dc.typeJournal articleen_US
dc.description.versionpublishedVersionen_US
dc.rights.holder© The Author(s). 2021en_US
dc.source.pagenumber15en_US
dc.source.volume22en_US
dc.source.journalGenome Biologyen_US
dc.identifier.doi10.1186/s13059-020-02231-9
dc.identifier.cristin1901666
dc.source.articlenumber6en_US
cristin.ispublishedtrue
cristin.fulltextoriginal
cristin.qualitycode2


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Navngivelse 4.0 Internasjonal
Except where otherwise noted, this item's license is described as Navngivelse 4.0 Internasjonal