James H. Eberwine, PhD

Co-Director of the PENN Genome Frontiers Institute,
Professor of Pharmacology
University of Pennsylvania, Philadelphia, PA

Emergent Properties of Single Cells as Defined by Single Cell Analyses

The analysis of single cell biology has provided unique and important insights into cellular function: For example, the relative and absolute abundances of cellular RNAs that comprise a cell’s expression profile are important for cellular function. Data will be presented showing that the expression profile of a cell acts as a phenotypic memory for cells, that when transferred between cells using our TIPeR technology enables the creation of cell types using investigator defined RNA populations and abundances.

In analyzing the transcriptomes of single cells for these studies we identified a novel class of RNAs that we call CIRTs (cytoplasmic intron retaining transcripts) that exist in all cells tested. The retained introns participate in multiple biological functions including subcellular targeting of RNA, exon selection and proper protein localization (all dependent upon cytoplasmic splicing). These data have compelled the elaboration of the Sentinel RNA hypothesis, which will be discussed.

To examine the role of RNA memory in vivo we have developed a procedure for characterization of the RNA complement from individual cells and subregions of cells that reside in their natural environment. Data from these experiments show that the microenvironment serves as an inhibitor of expression of a large numbers of genes and is important in generating RNA expression variability between cells. This innate microenvironment-associated transcriptome variability provides cells with their ability to tolerate their local phenotype and underlie all aspects of neuronal plasticity.

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