CRISPR/Cas9 mutagenesis of the Arabidopsis GROWTH-REGULATING FACTOR (GRF) gene family.

Autor:	Angulo J; Department of Plant Biology, University of Georgia, Athens, GA, United States., Astin CP; Division of Biology, University of Georgia, Athens, GA, United States., Bauer O; Department of Plant Biology, University of Georgia, Athens, GA, United States., Blash KJ; Division of Biology, University of Georgia, Athens, GA, United States., Bowen NM; Division of Biology, University of Georgia, Athens, GA, United States., Chukwudinma NJ; Division of Biology, University of Georgia, Athens, GA, United States., DiNofrio AS; Division of Biology, University of Georgia, Athens, GA, United States., Faletti DO; Division of Biology, University of Georgia, Athens, GA, United States., Ghulam AM; Division of Biology, University of Georgia, Athens, GA, United States., Gusinde-Duffy CM; Division of Biology, University of Georgia, Athens, GA, United States., Horace KJ; Division of Biology, University of Georgia, Athens, GA, United States., Ingram AM; Division of Biology, University of Georgia, Athens, GA, United States., Isaack KE; Division of Biology, University of Georgia, Athens, GA, United States., Jeong G; Division of Biology, University of Georgia, Athens, GA, United States., Kiser RJ; Division of Biology, University of Georgia, Athens, GA, United States., Kobylanski JS; Department of Plant Biology, University of Georgia, Athens, GA, United States., Long MR; Department of Plant Biology, University of Georgia, Athens, GA, United States., Manning GA; Department of Plant Biology, University of Georgia, Athens, GA, United States., Morales JM; Division of Biology, University of Georgia, Athens, GA, United States., Nguyen KH; Division of Biology, University of Georgia, Athens, GA, United States., Pham RT; Division of Biology, University of Georgia, Athens, GA, United States., Phillips MH; Department of Plant Biology, University of Georgia, Athens, GA, United States., Reel TW; Division of Biology, University of Georgia, Athens, GA, United States., Seo JE; Division of Biology, University of Georgia, Athens, GA, United States., Vo HD; Division of Biology, University of Georgia, Athens, GA, United States., Wukoson AM; Division of Biology, University of Georgia, Athens, GA, United States., Yeary KA; Department of Plant Biology, University of Georgia, Athens, GA, United States., Zheng GY; Department of Plant Biology, University of Georgia, Athens, GA, United States., Lukowitz W; Department of Plant Biology, University of Georgia, Athens, GA, United States.
Jazyk:	angličtina
Zdroj:	Frontiers in genome editing [Front Genome Ed] 2023 Oct 16; Vol. 5, pp. 1251557. Date of Electronic Publication: 2023 Oct 16 (Print Publication: 2023).
DOI:	10.3389/fgeed.2023.1251557
Abstrakt:	Genome editing in plants typically relies on T-DNA plasmids that are mobilized by Agrobacterium -mediated transformation to deliver the CRISPR/Cas machinery. Here, we introduce a series of CRISPR/Cas9 T-DNA vectors for minimal settings, such as teaching labs. Gene-specific targeting sequences can be inserted as annealed short oligonucleotides in a single straightforward cloning step. Fluorescent markers expressed in mature seeds enable reliable selection of transgenic or transgene-free individuals using a combination of inexpensive LED lamps and colored-glass alternative filters. Testing these tools on the Arabidopsis GROWTH-REGULATING FACTOR (GRF) genes, we were able to create a collection of predicted null mutations in all nine family members with little effort. We then explored the effects of simultaneously targeting two, four and eight GRF genes on the rate of induced mutations at each target locus. In our hands, multiplexing was associated with pronounced disparities: while mutation rates at some loci remained consistently high, mutation rates at other loci dropped dramatically with increasing number of single guide RNA species, thereby preventing a systematic mutagenesis of the family. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2023 Angulo, Astin, Bauer, Blash, Bowen, Chukwudinma, DiNofrio, Faletti, Ghulam, Gusinde-Duffy, Horace, Ingram, Isaack, Jeong, Kiser, Kobylanski, Long, Manning, Morales, Nguyen, Pham, Phillips, Reel, Seo, Vo, Wukoson, Yeary, Zheng and Lukowitz.)
Databáze:	MEDLINE
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