Targeted mutagenesis in the tetraploid switchgrass cultivar 'Alamo' using CRISPR-Cas9

dc.contributor.advisor Shui-zhang Fei Merrick, Paul
dc.contributor.department Horticulture 2018-08-11T11:52:10.000 2020-06-30T03:01:10Z 2020-06-30T03:01:10Z Fri Jan 01 00:00:00 UTC 2016 2001-01-01 2016-01-01
dc.description.abstract <p>Switchgrass (Panicum virgatum L.) is a highly self-incompatible perennial grass species which has been designated as a model bioenergy crop by the U.S. Department of Energy. To enhance breeding efforts of switchgrass we tested the efficacy of the targeted gene editing tool CRISPR-Cas9 in the tetraploid cultivar ‘Alamo’. Agrobacterium mediated transformation of embryogenic callus tissue averaged a transformation efficiency of 12.0% across experiments targeting four genes: Flowering Locus T (FT), APETALA 1 (AP1), Brassinosteroid Receptor Insensitive 1 (BRI1), and Phosphoglycerate Mutase (PGM). Out of 77 PCR confirmed transgenic events, a single event contained a heterozygous, CRISPR-Cas9 mediated 1 base pair deletion in the PGM target gene. Transgene silencing due to switchgrass’s highly methylated genome may play a role in the low mutation frequency. The mutation was obtained using a single sgRNA, however multiplex CRISPR using 2 sgRNA’s as well as tRNA constructs were also tested.</p>
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dc.identifier archive/
dc.identifier.articleid 6070
dc.identifier.contextkey 8882605
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath etd/15063
dc.language.iso en
dc.source.bitstream archive/|||Fri Jan 14 20:35:09 UTC 2022
dc.subject.disciplines Genetics
dc.subject.keywords Genetics
dc.title Targeted mutagenesis in the tetraploid switchgrass cultivar 'Alamo' using CRISPR-Cas9
dc.type article
dc.type.genre thesis
dspace.entity.type Publication
relation.isOrgUnitOfPublication df043cd4-424c-49f5-8685-318972aae642 Genetics thesis Master of Science
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