CRISPR-based technologies for studying RNA

CRISPR-Cas9 has transformed our ability to study DNA by manipulating genomic information in a programmable fashion. The RNA-targeting CRISPR effector protein Cas13 is poised to revolutionize the study of RNA in a similar fashion, with numerous applications in RNA knockdown, detection, editing, and imaging. To fully realize the potential of Cas13 as a platform technology to detect and manipulate RNA, we need a better understanding of how target sequence and structure govern Cas13’s ability to detect and cleave RNA. To address this key challenge, my lab at Princeton is developing a diverse set of programmable Cas13 tools and technologies for studying RNA in vitro, in cells, and in vivo. These include technologies for detecting viruses and bacterial pathogens, and for profiling the immune response to infection. We also are developing technologies for RNA imaging and perturbation in eukaryotic cells. In addition, we are investigating the molecular mechanisms by which Cas13 can target RNA so that we can further enhance its programmable RNA-targeting properties. We are applying these technologies to answer an array of biological questions that have major implications for human health. Our interests span infectious diseases and human genetic diseases, many of which are associated with RNA dysregulation or mutations in RNA-binding proteins.

Learning Objectives:

1. Describe the role of Cas13 in RNA-targeting compared to Cas9’s role in DNA editing.

2. Explain how Cas13 can be applied for RNA knockdown, detection, editing, and imaging.

3. Identify factors such as sequence and structure that influence Cas13’s ability to cleave RNA.