Characterization of Programmable Transcription Activators in the Model Monocot Setaria viridis Via Protoplast Transfection

Adam Sychla, Juan A Casas Mollano, Matthew H Zinselmeier, Michael Smanski

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


Recent advances in DNA synthesis and assembly allow for genetic constructs to be designed and constructed in high throughput. Characterizing large numbers of variant genetic designs is not feasible with low-throughput and time-consuming plant transformation workflows. Protoplast transformation offers a rapid, high-throughput compatible alternative for testing genetic constructs in plant-relevant molecular environments. Here, we describe a protocol for protoplast transformation using a recent experiment in genetic optimization of dCas9-based programmable transcription activators as an example.

Original languageEnglish (US)
Pages (from-to)223-244
Number of pages22
JournalMethods in molecular biology (Clifton, N.J.)
StatePublished - 2022

Bibliographical note

Funding Information:
AS and MZ were funded by NIGMS Biotechnology Training grant NIHT32GM008347. MJS and JACM are part of a team supported by the Advanced Plant Technologies program, DARPA Award HR001118C0146. MJS and MZ are supported by USDA NIFA award 2018-33522-28747.

Publisher Copyright:
© 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.


  • High throughput
  • Monocot
  • Protoplasts
  • Setaria viridis
  • Transcription
  • Transfection
  • dCas9

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.


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