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US 2021/0172017 A1Published application
Methods of in vivo evaluation of gene function
AssigneesThe Broad Institute, Inc., Massachusetts Institute of Technology, President and Fellows of Harvard College
InventorsXin Jin, Paola Arlotta, Aviv Regev, Feng Zhang, Sean Simmons
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Abstract
Described herein are methods and uses thereof for in vivo evaluating functions of multiple genes in parallel by combining in utero genetic perturbation of progenitor cells and single-cell transcriptomic profiling of progeny cells in animals. These methods can be used, among other things, to reveal in vivo gene functions in a cell type-specific manner.
Claims28
A method of identifying functions of a plurality of genes in parallel in vivo, comprising:
- a. introducing, in vivo, a plurality of genetic perturbations in each of a plurality of progenitor cells in a Cas animal model, wherein each genetic perturbation is operatively coupled to a reporter gene and a barcode;
- b. generating an enriched perturbed cell population by enriching for cells expressing the reporter;
- c. identifying cell types and corresponding perturbations via scRNA-seq in the enriched perturbed cell population;
- and
- d. detecting one or more gene modules that co-vary within a cell type in the enriched perturbed cell population.
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