in situ hybridization (ISH) has become the mainstay of many approaches to understanding gene expression in biology and pathology, whereby labelled probes complementary to mRNA sequences – i.e., anti-sense probes – are used to localize mRNAs in cells and tissue sections. In existence for some 5 decades, the basic ISH technique still requires substantial optimization covering a range of skills, from basic molecular biology to tissue isolation and preparation and is subject to further variability amongst operators. RNAscope offers a superior alternative and scalable approach to classical ISH, with recent iterations allowing resolution of much shorter targets and even junctional sequences. This webcast provides an introduction to this comparatively new commercial technology and its use in answering questions previously addressed by classical techniques. It is presented from the viewpoint of an ISH user with 30-years of classical experience who has switched to RNAscope and BaseScope, with a particular emphasis on its use in studying the nervous system. As a tangible example, the use of RNAscope was instrumental in validating “hits” identified from interrogating single-neuron transcriptomes obtained by sequencing single neuronal nuclei of the human brain (Science 352, 1586-1590 (2016)), and examples from this and other research efforts will be presented.

In this webcast you will learn; advances in in situ hybridization and RNAscope enable RNA gene expression analysis off-the-shelf and single day assays BaseScope gene expression analysis enables splice variance and short target analysis mRNA analysis in neuroscience research where no antibodies are available; detection of long non-coding RNA (lncRNA) RNA-seq to target validation and validation of (cell type-specific) genetic modifications; and ability to visualize in situ expression of virtually any gene, any species and any tissue. You will also have the opportunity to ask questions of our speaker, live during the broadcast!