Method for RNA extraction and transcriptomic analysis of single fungal spores

• Main Authors: Ivey A. Geoghegan, Richard D. Emes, David B. Archer, Simon V. Avery
• Format: Article
• Language: English
• Published: Elsevier 2020-01-01
• Series: MethodsX
• Subjects: RNA extraction; Single-cell transcriptomics; Conidia; Spores; Aspergillus niger; Phenotypic heterogeneity
• Online Access: http://www.sciencedirect.com/science/article/pii/S221501611930336X

Transcriptomic analysis of single cells has been increasingly in demand in recent years, thanks to technological and methodological advances as well as growing recognition of the importance of individuals in biological systems. However, the majority of these studies have been performed in mammalian cells, due to their ease of lysis and high RNA content. No single cell transcriptomic analysis has yet been described in microbial spores, even though it is known that heterogeneity at the phenotype level exists among individual spores. Transcriptomic analysis of single spores is challenging, in part due to the physically robust nature of the spore wall. This precludes the use of methods commonly used for mammalian cells. Here, we describe a simple method for extraction and amplification of transcripts from single fungal conidia (asexual spores), and its application in single-cell transcriptomics studies. The method can also be used for studies of small numbers of fungal conidia, which may be necessary in the case of limited sample availability, low-abundance transcripts or interest in small subpopulations of conidia. • The method allows detection of transcripts from single conidia of Aspergillus niger • The method allows detection of genomic DNA from single conidia of Aspergillus niger