Massachusetts Institute of Technology

The Broad Institute, Inc.

Jacob Borrajo

Paul Blainey

Atray Dixit

Mohamad Najia

This disclosure provides compositions and methods of using these compositions to mediate a targeted trans-splicing event on a pre-mRNA in a cell.

Targeted trans-splicing using CRISPR/Cas13

The present invention provides for methods to obtain multiple information-rich samples at different time points from the same cell while minimally disrupting the cell. The subject matter disclosed herein is generally related to nucleic acid constructs for continuous monitoring of live cells. Specifically, the subject matter disclosed herein is directed to nucleic acid constructs that encode a fusion protein and a construct RNA sequence that induce live cells to self-report cellular contents while maintaining cell viability. The present invention may be used to monitor gene expression in single cells while maintaining cell viability.

Constructs for continuous monitoring of live cells

Embodiments disclosed herein provide nucleic acid constructs and methods of use thereof that induce a live cell to give off sub-samples of the cell's cytosolic content. The tem 'cell' as used herein may be any cell type. In certain example embodiments, the cells are mammalian cells. The sampling can be general or can be targeted to a particular class of molecules or to specific types of molecules. The constructs facilitate generation of a read-out for high-throughput screens by combining engineered export with simple bulk sample and sample processing. Live cell sampling enables time course measurements and expands, for example, the applicability of transcriptional profiles obtained by single cell gene expression analysis.

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Cambridge, MA, United States of America

Jacob Borrajo