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

dc.contributor.advisor	Shui-zhang Fei
dc.contributor.author	Merrick, Paul
dc.contributor.department	Horticulture
dc.date	2018-08-11T11:52:10.000
dc.date.accessioned	2020-06-30T03:01:10Z
dc.date.available	2020-06-30T03:01:10Z
dc.date.copyright	Fri Jan 01 00:00:00 UTC 2016
dc.date.embargo	2001-01-01
dc.date.issued	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>
dc.format.mimetype	application/pdf
dc.identifier	archive/lib.dr.iastate.edu/etd/15063/
dc.identifier.articleid	6070
dc.identifier.contextkey	8882605
dc.identifier.doi	https://doi.org/10.31274/etd-180810-4666
dc.identifier.s3bucket	isulib-bepress-aws-west
dc.identifier.submissionpath	etd/15063
dc.identifier.uri	https://dr.lib.iastate.edu/handle/20.500.12876/29247
dc.language.iso	en
dc.source.bitstream	archive/lib.dr.iastate.edu/etd/15063/Merrick_iastate_0097M_15630.pdf\|\|\|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
thesis.degree.discipline	Genetics
thesis.degree.level	thesis
thesis.degree.name	Master of Science

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