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Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies
OBJECTIVES: Sorghum is one of the most recalcitrant species for transformation. Considering the time and effort required for stable transformation in sorghum, establishing a transient system to screen the efficiency and full functionality of vector constructs is highly desirable. RESULTS: Here, we r...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7045639/ https://www.ncbi.nlm.nih.gov/pubmed/32103777 http://dx.doi.org/10.1186/s13104-020-04968-9 |
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author | Sharma, Rita Liang, Yan Lee, Mi Yeon Pidatala, Venkataramana R. Mortimer, Jenny C. Scheller, Henrik V. |
author_facet | Sharma, Rita Liang, Yan Lee, Mi Yeon Pidatala, Venkataramana R. Mortimer, Jenny C. Scheller, Henrik V. |
author_sort | Sharma, Rita |
collection | PubMed |
description | OBJECTIVES: Sorghum is one of the most recalcitrant species for transformation. Considering the time and effort required for stable transformation in sorghum, establishing a transient system to screen the efficiency and full functionality of vector constructs is highly desirable. RESULTS: Here, we report an Agrobacterium-mediated transient transformation assay with intact sorghum leaves using green fluorescent protein as marker. It also provides a good monocot alternative to tobacco and protoplast assays with a direct, native and more reliable system for testing single guide RNA (sgRNA) expression construct efficiency. Given the simplicity and ease of transformation, high reproducibility, and ability to test large constructs, this method can be widely adopted to speed up functional genomic and genome editing studies. |
format | Online Article Text |
id | pubmed-7045639 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-70456392020-03-03 Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies Sharma, Rita Liang, Yan Lee, Mi Yeon Pidatala, Venkataramana R. Mortimer, Jenny C. Scheller, Henrik V. BMC Res Notes Research Note OBJECTIVES: Sorghum is one of the most recalcitrant species for transformation. Considering the time and effort required for stable transformation in sorghum, establishing a transient system to screen the efficiency and full functionality of vector constructs is highly desirable. RESULTS: Here, we report an Agrobacterium-mediated transient transformation assay with intact sorghum leaves using green fluorescent protein as marker. It also provides a good monocot alternative to tobacco and protoplast assays with a direct, native and more reliable system for testing single guide RNA (sgRNA) expression construct efficiency. Given the simplicity and ease of transformation, high reproducibility, and ability to test large constructs, this method can be widely adopted to speed up functional genomic and genome editing studies. BioMed Central 2020-02-27 /pmc/articles/PMC7045639/ /pubmed/32103777 http://dx.doi.org/10.1186/s13104-020-04968-9 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Note Sharma, Rita Liang, Yan Lee, Mi Yeon Pidatala, Venkataramana R. Mortimer, Jenny C. Scheller, Henrik V. Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies |
title | Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies |
title_full | Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies |
title_fullStr | Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies |
title_full_unstemmed | Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies |
title_short | Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies |
title_sort | agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies |
topic | Research Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7045639/ https://www.ncbi.nlm.nih.gov/pubmed/32103777 http://dx.doi.org/10.1186/s13104-020-04968-9 |
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