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

doi:10.1007/978-1-0716-2164-6_16

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 journal › Article › peer-review

2 Scopus citations

Abstract

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 language	English (US)
Pages (from-to)	223-244
Number of pages	22
Journal	Methods in molecular biology (Clifton, N.J.)
Volume	2464
DOIs	https://doi.org/10.1007/978-1-0716-2164-6_16
State	Published - 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.

Keywords

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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10.1007/978-1-0716-2164-6_16

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abstract = "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.",

keywords = "High throughput, Monocot, Protoplasts, Setaria viridis, Transcription, Transfection, dCas9",

author = "Adam Sychla and {Casas Mollano}, {Juan A} and Zinselmeier, {Matthew H} and Michael Smanski",

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: {\textcopyright} 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",

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AU - Zinselmeier, Matthew H

AU - Smanski, Michael

N1 - 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.

PY - 2022

Y1 - 2022

N2 - 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.

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KW - High throughput

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KW - Protoplasts

KW - Setaria viridis

KW - Transcription

KW - Transfection

KW - dCas9

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

Abstract

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Keywords

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