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dc.contributor.author
Andersson, Mariette
dc.contributor.author
Turesson, Helle
dc.contributor.author
Olsson, Niklas
dc.contributor.author
Fält, Ann Sofie
dc.contributor.author
Ohlsson, Pia
dc.contributor.author
Gonzalez, Matías Nicolás
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dc.contributor.author
Samuelsson, Mathias
dc.contributor.author
Hofvander, Per
dc.date.available
2020-03-25T18:52:11Z
dc.date.issued
2018-12
dc.identifier.citation
Andersson, Mariette; Turesson, Helle; Olsson, Niklas; Fält, Ann Sofie; Ohlsson, Pia; et al.; Genome editing in potato via CRISPR-Cas9 ribonucleoprotein delivery; Wiley Blackwell Publishing, Inc; Physiologia Plantarum; 164; 4; 12-2018; 378-384
dc.identifier.issn
0031-9317
dc.identifier.uri
http://hdl.handle.net/11336/100759
dc.description.abstract
Clustered regularly interspaced short palindromic repeats and CRISPR-associated protein-9 (CRISPR-Cas9) can be used as an efficient tool for genome editing in potato (Solanum tuberosum). From both a scientific and a regulatory perspective, it is beneficial if integration of DNA in the potato genome is avoided. We have implemented a DNA-free genome editing method, using delivery of CRISPR-Cas9 ribonucleoproteins (RNPs) to potato protoplasts, by targeting the gene encoding a granule bound starch synthase (GBSS, EC 2.4.1.242). The RNP method was directly implemented using previously developed protoplast isolation, transfection and regeneration protocols without further adjustments. Cas9 protein was preassembled with RNA produced either synthetically or by in vitro transcription. RNP with synthetically produced RNA (cr-RNP) induced mutations, i.e. indels, at a frequency of up to 9%, with all mutated lines being transgene-free. A mutagenesis frequency of 25% of all regenerated shoots was found when using RNP with in vitro transcriptionally produced RNA (IVT–RNP). However, more than 80% of the shoots with confirmed mutations had unintended inserts in the cut site, which was in the same range as when using DNA delivery. The inserts originated both from DNA template remnants from the in vitro transcription, and from chromosomal potato DNA. In 2–3% of the regenerated shoots from the RNP-experiments, mutations were induced in all four alleles resulting in a complete knockout of the GBSS enzyme function.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Wiley Blackwell Publishing, Inc
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dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by/2.5/ar/
dc.subject
POTATO
dc.subject
STARCH
dc.subject
TRANSGENE FREE
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CRISPR CAS9
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PROTOPLASTS
dc.subject.classification
Otras Ciencias Biológicas
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dc.subject.classification
Ciencias Biológicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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dc.title
Genome editing in potato via CRISPR-Cas9 ribonucleoprotein delivery
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2020-03-25T14:01:47Z
dc.journal.volume
164
dc.journal.number
4
dc.journal.pagination
378-384
dc.journal.pais
Reino Unido
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dc.journal.ciudad
Londres
dc.description.fil
Fil: Andersson, Mariette. Swedish University Of Agricultural Sciences; Suecia
dc.description.fil
Fil: Turesson, Helle. Swedish University Of Agricultural Sciences; Suecia
dc.description.fil
Fil: Olsson, Niklas. Swedish University Of Agricultural Sciences; Suecia
dc.description.fil
Fil: Fält, Ann Sofie. Swedish University Of Agricultural Sciences; Suecia
dc.description.fil
Fil: Ohlsson, Pia. Swedish University Of Agricultural Sciences; Suecia
dc.description.fil
Fil: Gonzalez, Matías Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; Argentina
dc.description.fil
Fil: Samuelsson, Mathias. Lyckeby Starch AB; Suecia
dc.description.fil
Fil: Hofvander, Per. Swedish University Of Agricultural Sciences; Suecia
dc.journal.title
Physiologia Plantarum
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dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://doi.wiley.com/10.1111/ppl.12731
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1111/ppl.12731
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