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Convergence of distinct RNA-silencing pathways on GW182/Tnrc6 [sgRNA screen]

ABSTRACT: sgRNA screens for factors influencing Trim71 silencing activity.

ORGANISM(S): Mus musculus

PROVIDER: GSE214708 | GEO | 2023/06/26

REPOSITORIES: GEO

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Convergence of distinct RNA-silencing pathways on GW182/Tnrc6

Project description:The RNA-binding protein Trim71/Lin41 is a phylogenetically conserved developmental regulator that functions in mammalian stem cell reprogramming, brain development and cancer. Trim71 recognizes target mRNAs through hairpin motifs and silences them through molecular mechanisms that await identification. Deposited data are mass spectrometry data from immunoprecipitation experiments with endogenously tagged Trim71, Ago2, and Tnrc6a. Experiments are carried out in mouse embryonic stem cells (mESCs) and uncover reciprocal interactions of Trim71, Ago2, and Tnrc6a. Trim71 protein interactions are largely independent of Ago2 levels, but strongly depend on the presence of RNA. A second set of experiments is mass spectrometry data from immunoprecipitation experiments in mESCs overexpressing a tagged peptide derived from Homo sapiens TNRC6B (‘FLAG-HA::T6BWT::Cherry’), that is known to block the Tnrc6-Ago2 interaction. We show that a wild-type version strongly binds Ago1 and Ago2, but not Trim71, while a mutant version binds neither Ago1, 2, nor Trim71.

2023-07-04 | PXD034537 | Pride

Convergence of distinct RNA-silencing pathways on GW182/Tnrc6 II

Project description:RNA-sequencing in mouse embryonic stem cells: Ctrl: GFP_5xT71BS (mES [129S6/SvEvTac], yGlob [GFP_5xT71BS]. Trim71-/-: GFP_5xT71BS (mES [129S6/SvEvTac], yGlob [GFP_5xT71BS] Trim71 -/-. Note pervasive gene expression changes in samples with TNRC6A overexpression in comparison with samples with CAS9 overexpression.

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Convergence of distinct RNA-silencing pathways on GW182/Tnrc6 [RNA-seq]

Project description:RNA-sequencing in mouse embryonic stem cells. Experiment 1 (Samples 1-9): Base cell line (WT): GFP_5xT71BS (mES [129S6/SvEvTac], yGlob [GFP_5xT71BS], stable transfection FLAG-Cas9::T2A::Blasticidin). Trim71 KO: GFP_5xT71BS Trim71-/-. Ago2 KO: GFP_5xT71BS Ago2-/-. Experiment 2 (Samples 10-21): parental cell line: GFP_5xT71BS. T6B_Wt: Overexpression of T6B_WT, T6B_Mut: Overexpression of T6B_MUT. Trim71_KO: GFP_5xT71BS, Trim71-/- transfected with empty plasmid. Ago2_KO: GFP_5xT71BS, Ago2-/- transfected with empty plasmid.

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Convergence of distinct RNA-silencing pathways on GW182/Tnrc6

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Convergence of distinct RNA-silencing pathways on GW182/Tnrc6 [RIP-seq]

Project description:RIP-sequencing in mouse embryonic stem cells. Experiment 1 (Samples 1-18): Base cell line (untagged control cells): GFP_5xT71BS (mES [129S6/SvEvTac], yGlob [GFP_5xT71BS], stable transfection FLAG-Cas9::T2A::Blasticidin). HA-Tnrc6a: endogenously HA-AVI-tagged Tnrc6a. HA-Tnrc6a_Trim71KO: HA-AVI-tagged Tnrc6a, Trim71-/-. Experiment 2 (Samples 19-42): Ctrl: base cell line GFP_5xT71BS. Tnrc6a: endogenously HA-AVI-tagged Tnrc6a. Tnrc6a_Trim71KO: HA-AVI-tagged Tnrc6a, Trim71-/-. Tnrc6a_Ago2KO: HA-AVI-tagged Tnrc6a, Ago2-/-.

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Convergence of distinct RNA-silencing pathways on GW182/Tnrc6 [sgRNA screen]

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Convergence of distinct RNA-silencing pathways on GW182/Tnrc6 [RIP-seq II]

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Convergence of distinct RNA-silencing pathways on GW182/Tnrc6

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CRISPR activation screen identifies BCL-2 proteins and B3GNT2 as drivers of cancer resistance to T cell-mediated cytotoxicity (sgRNA distribution)

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