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农杆菌介导的高粱叶片瞬时转化用于加速功能基因组学和基因组编辑研究。

Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies.

作者信息

Sharma Rita, Liang Yan, Lee Mi Yeon, Pidatala Venkataramana R, Mortimer Jenny C, Scheller Henrik V

机构信息

Joint BioEnergy Institute, Emeryville, CA, 94608, USA.

Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA.

出版信息

BMC Res Notes. 2020 Feb 27;13(1):116. doi: 10.1186/s13104-020-04968-9.

Abstract

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.

摘要

目的

高粱是最难转化的物种之一。鉴于高粱稳定转化所需的时间和精力,建立一个用于筛选载体构建体效率和完整功能的瞬时系统是非常必要的。

结果

在此,我们报告了一种以绿色荧光蛋白为标记物,利用完整高粱叶片进行农杆菌介导的瞬时转化分析方法。它还为烟草和原生质体分析提供了一个很好的单子叶替代方法,是一种直接、天然且更可靠的测试单导向RNA(sgRNA)表达构建体效率的系统。鉴于该方法转化简单易行、重复性高且能够测试大型构建体,它可被广泛采用以加速功能基因组学和基因组编辑研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ffc4/7045639/fc37c71dfefd/13104_2020_4968_Fig1_HTML.jpg

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