title: Manipulation of Oat Protoplasts for Transient Expression Assays.
abstract: Oat protoplasts are a useful and convenient system to study transient expression using whole cells. Nucleic acid can rapidly be introduced into live cells, and, depending on the assay, results can be collected the same day. Compared to plant tissue, oat cell suspension cultures provide a simple, high yielding, and consistent means to isolate protoplasts. Here, we describe how to generate an oat cell suspension culture from callus grown on solidified medium, and how to maintain the oat cells in cell suspension culture for protoplast preparation. Following the culturing procedure, we describe how to isolate oat protoplasts from cell suspension culture by enzymatic digestion of the cell walls and to transiently express nucleic acid (DNA or RNA) into the cells by electroporation.
authors.affiliation: University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Robyn
authors.initials: R
authors.lastname: Roberts
authors.affiliation: University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Jincan
authors.initials: J
authors.lastname: Zhang
authors.affiliation: University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Nicole
authors.initials: N
authors.lastname: Mihelich
authors.affiliation: University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Danielle
authors.initials: D
authors.lastname: Savino
authors.affiliation: University of Wisconsin-Madison, Madison, WI, USA. rakotondrafa@wisc.edu.
authors.forename: Aurélie M
authors.initials: AM
authors.lastname: Rakotondrafara
mesh_terms: D018554:Avena / Q000235:genetics*; D018929:Cell Culture Techniques; D018274:Electroporation; D015870:Gene Expression; D011523:Protoplasts; D014162:Transfection
keywords: Cell callus; Electroporation; Nucleic acid; Oat cell suspension culture; Oat protoplast; Transient expression