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dc.contributor.authorKim, Dongjin
dc.contributor.authorAlptekin, Burcu
dc.contributor.authorBudak, Hikmet
dc.date.accessioned2018-04-24T18:48:23Z
dc.date.available2018-04-24T18:48:23Z
dc.date.issued2017-09
dc.identifier.citationKim, Dongjin, Burcu Alptekin, and Hikmet Budak. "CRISPR/Cas9 genome editing in wheat." Functional and Integrative Genomics 18, no. 1 (September 2017): 31-41. DOI: 10.1007/s10142-017-0572-x.en_US
dc.identifier.issn1438-793X
dc.identifier.urihttps://scholarworks.montana.edu/xmlui/handle/1/14505
dc.description.abstractGenome editing has been a long-term challenge for molecular biology research, particularly for plants possess complex genome. The recently discovered Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a versatile tool for genome editing which enables editing of multiple genes based on the guidance of small RNAs. Even though the efficiency of CRISPR/Cas9 system has been shown with several studies from diploid plants, its application remains a challenge for plants with polyploid and complex genome. Here, we applied CRISPR/Cas9 genome editing system in wheat protoplast to conduct the targeted editing of stress-responsive transcription factor genes, wheat dehydration responsive element binding protein 2 (TaDREB2) and wheat ethylene responsive factor 3 (TaERF3). Targeted genome editing of TaDREB2 and TaERF3 was achieved with transient expression of small guide RNA and Cas9 protein in wheat protoplast. The effectiveness of mutagenesis in wheat protoplast was confirmed with restriction enzyme digestion assay, T7 endonuclease assay, and sequencing. Furthermore, several off-target regions for designed sgRNAs were analyzed, and the specificity of genome editing was confirmed with amplicon sequencing. Overall results suggested that CRISPR/Cas9 genome editing system can easily be established on wheat protoplast and it has a huge potentiality for targeted manipulation of wheat genome for crop improvement purposes.en_US
dc.description.sponsorshipMontana Wheat and Barley Committee Grant #MDA/MWBC CY5416-462; Winifred-Asbjornson Plant Science Endowment;en_US
dc.titleCRISPR/Cas9 genome editing in wheaten_US
dc.typeArticleen_US
mus.citation.extentfirstpage31en_US
mus.citation.extentlastpage41en_US
mus.citation.issue1en_US
mus.citation.journaltitleFunctional and Integrative Genomicsen_US
mus.citation.volume18en_US
mus.identifier.categoryLife Sciences & Earth Sciencesen_US
mus.identifier.doi10.1007/s10142-017-0572-xen_US
mus.relation.collegeCollege of Agricultureen_US
mus.relation.departmentPlant Sciences & Plant Pathology.en_US
mus.relation.universityMontana State University - Bozemanen_US
mus.data.thumbpage5en_US
mus.contributor.orcidAlptekin, Burcu|0000-0002-0041-0240en_US
mus.contributor.orcidBudak, Hikmet|0000-0002-2556-2478en_US
mus.contributor.orcidKim, Dongjin|0000-0003-4175-9817en_US


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