利用水稻原生质体在大规模平台上进行高效瞬时基因转染以进行转录活性分析。

High-Efficient Transient Gene Transfection for Transcriptional Activity Assay Using Rice Protoplasts on Large-Scale Platform.

机构信息

Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan.

Global Zero-Emission Research Center, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan.

出版信息

Methods Mol Biol. 2025;2869:183-192. doi: 10.1007/978-1-0716-4204-7_18.

DOI:10.1007/978-1-0716-4204-7_18

PMID:39499478

Abstract

Transient gene expression in protoplasts offers a straightforward and efficient approach for investigating gene function. Here, I introduce a method that utilizes a 96-well plate format to assess transcriptional activation capacity of transcription factors through transient expression system in rice protoplasts. This method includes plant cultivation, protoplast preparation, transfection, and transactivation assay and allows for the simultaneous measurement and validation of 23 samples with 4 replicates per sample at a time.

摘要

原生质体中的瞬时基因表达为研究基因功能提供了一种直接有效的方法。在这里，我介绍了一种利用 96 孔板格式通过水稻原生质体中的瞬时表达系统评估转录因子转录激活能力的方法。该方法包括植物培养、原生质体制备、转染和反式激活测定，可同时测量和验证每个样品的 4 个重复 23 个样品。