Whole-mount smFISH allows combining RNA and protein quantification at cellular and subcellular resolution.

ABSTRACT: Multicellular organisms result from complex developmental processes largely orchestrated through the quantitative spatiotemporal regulation of gene expression. Yet, obtaining absolute counts of messenger RNAs at a three-dimensional resolution remains challenging, especially in plants, owing to high levels of tissue autofluorescence that prevent the detection of diffraction-limited fluorescent spots. In situ hybridization methods based on amplification cycles have recently emerged, but they are laborious and often lead to quantification biases. In this article, we present a simple method based on single-molecule RNA fluorescence in situ hybridization to visualize and count the number of mRNA molecules in several intact plant tissues. In addition, with the use of fluorescent protein reporter

SUBMITTER: Zhao L 

PROVIDER: S-EPMC10356603 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature