High-throughput Flow Cytometry Assay to Investigate TDP43 Splicing Function

被引：2

作者：

Schmidt, H. Broder ^{[1
]}

Rohatgi, Rajat ^{[1
,2
,3
]}

机构：

[1] Stanford Univ, Dept Biochem, Sch Med, Stanford, CA 94305 USA

[2] Stanford Univ, Sch Med, Dept Med, Stanford, CA 94305 USA

[3] Stanford Univ, Stanford Canc Inst, Sch Med, Stanford, CA USA

来源：

BIO-PROTOCOL | 2020年 / 10卷 / 08期

基金：

美国国家卫生研究院;

关键词：

RNA splicing; Minigene; Flow cytometry; TDP43; Amyotrophic lateral sclerosis; TDP-43; PROTEINS; DISEASE;

D O I：

10.21769/BioProtoc.3594

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Mutations in RNA-binding proteins (RBPs) such as TDP43 are associated with transcriptome-wide splicing defects and cause severe neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). The impact of RBP mutations on splicing function is routinely studied using PCR-based bulk measurements. However, the qualitative and low-throughput nature of this assay make quantitative and systematic analyses, as well as screening approaches, difficult to implement. To overcome this hurdle, we have developed a quantitative, high-throughput flow cytometry assay to investigate TDP43 splicing function on a single-cell level

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页数：11

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