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In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Mus musculus

PROVIDER: GSE183502 | GEO | 2022/01/23

REPOSITORIES: GEO

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In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration [liver-RNA-seq]

Project description:Identifying new pathways that regulate mammalian regeneration is challenging due to the paucity of in vivo screening approaches. We employed pooled CRISPR knockout and activation screening in the regenerating liver to evaluate 164 chromatin regulatory proteins. Both screens identified imitation-SWI chromatin remodeling components Baz2a and Baz2b, not previously implicated in regeneration. In vivo sgRNA, siRNA, and knockout strategies against either paralog confirmed increased regeneration. Distinct BAZ2-specific bromodomain inhibitors GSK2801 and BAZ2-ICR resulted in accelerated liver healing after diverse injuries. Inhibitor treated mice also exhibited improved regeneration in an inflammatory bowel disease model, suggesting multi-tissue applicability. Transcriptomics on regenerating livers showed increases in ribosomal and cell cycle mRNAs. Surprisingly, CRISPRa screening to define mechanisms showed that overproducing ribosomal proteins was sufficient to drive regeneration, while Rpl24 haploinsufficiency was rate limiting for BAZ2 inhibition mediated regeneration. The discovery of regenerative roles for imitation-SWI components provides immediate strategies to enhance tissue repair.

2022-01-25 | GSE183501 | GEO

In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration [H2-ChIP-seq]

2022-01-25 | GSE183500 | GEO

In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration [liver-ChIP-seq]

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In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration [H2-ATAC-seq]

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In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration

Project description:In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration

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In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration [H2-ChIP-seq]

Project description:In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration [H2-ChIP-seq]

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In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration [H2-ATAC-seq]

Project description:In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration [H2-ATAC-seq]

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In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration [liver-RNA-seq]

Project description:In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration [liver-RNA-seq]

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In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration [liver-ChIP-seq]

Project description:In vivo CRISPR screening reveals druggable regulators of mammalian tissue regeneration [liver-ChIP-seq]

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Imaging-based pooled CRISPR screening reveals regulators of lncRNA localization.

Project description:Pooled-library CRISPR screening provides a powerful means to discover genetic factors involved in cellular processes in a high-throughput manner. However, the phenotypes accessible to pooled-library screening are limited. Complex phenotypes, such as cellular morphology and subcellular molecular organization, as well as their dynamics, require imaging-based readout and are currently beyond the reach of pooled-library CRISPR screening. Here we report an all imaging-based pooled-library CRISPR screening approach that combines high-content phenotype imaging with high-throughput single guide RNA (sgRNA) identification in individual cells. In this approach, sgRNAs are codelivered to cells with corresponding barcodes placed at the 3' untranslated region of a reporter gene using a lentiviral delivery system with reduced recombination-induced sgRNA-barcode mispairing. Multiplexed error-robust fluorescence in situ hybridization (MERFISH) is used to read out the barcodes and hence identify the sgRNAs with high accuracy. We used this approach to screen 162 sgRNAs targeting 54 RNA-binding proteins for their effects on RNA localization to nuclear compartments and uncovered previously unknown regulatory factors for nuclear RNA localization. Notably, our screen revealed both positive and negative regulators for the nuclear speckle localization of a long noncoding RNA, MALAT1, suggesting a dynamic regulation of lncRNA localization in subcellular compartments.

| S-EPMC6561216 | biostudies-literature

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