Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

RNA-Seq of mouse embryonic stem cells after knockout and re-introduction of Sall4a

ABSTRACT: Sall4 is a stem cell factor which is important for embryogenesis. In order to address transcriptional changes, we re-introduce Sall4 in homozygous Sall4 knockout (KO) mouse embryonic stem cells through random integration of Sall4 cDNA using a Piggybac vector which is Dox inducible. We also re-introduce EGFP in homozygous Sall4 knockout (KO) mouse embryonic stem cells and use it as a negative control.

INSTRUMENT(S): HiSeq X Five

ORGANISM(S): Mus musculus

SUBMITTER: Raphaël Pantier

PROVIDER: E-MTAB-9202 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

SALL4 controls cell fate in response to DNA base composition.

Pantier Raphaël R Chhatbar Kashyap K Quante Timo T Skourti-Stathaki Konstantina K Cholewa-Waclaw Justyna J Alston Grace G Alexander-Howden Beatrice B Lee Heng Yang HY Cook Atlanta G AG Spruijt Cornelia G CG Vermeulen Michiel M Selfridge Jim J Bird Adrian A

Molecular cell 20210105 4

Mammalian genomes contain long domains with distinct average compositions of A/T versus G/C base pairs. In a screen for proteins that might interpret base composition by binding to AT-rich motifs, we identified the stem cell factor SALL4, which contains multiple zinc fingers. Mutation of the domain responsible for AT binding drastically reduced SALL4 genome occupancy and prematurely upregulated genes in proportion to their AT content. Inactivation of this single AT-binding zinc-finger cluster mi ...[more]

PMID: 33406384

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Project description:Increasing studies suggest that SALL4 may play vital roles in leukemogenesis. We have used chromatin immunoprecipitation followed by microarray hybridization as a screening tool to determine potential genes that may account for the role SALL4 plays in leukemogenesis. Analysis of SALL4 binding sites reveals that genes involved in cell death, cancer, DNA replication/repair, and cell cycle were highly enriched (p<0.05). These genes include 38 important apoptosis-inducing genes (TNF, TP53, PTEN, CARD9, CARD11, CYCS, LTA) and apoptosis-inhibiting genes (Bmi-1, BCL2, XIAP, DAD1, TEGT). Real-time PCR has shown that expression levels of these genes changed significantly after SALL4 knockdown, which ubiquitously led to cell apoptosis. Flow cytometry revealed that reduction of SALL4 expression in NB4 and other leukemia cell lines dramatically increased caspase-3, Annexin V, and DNA fragmentation activity. Bromodeoxyuridine-incorporation assays showed decreased numbers of S phase cells and increased numbers of G1- and G2- phase cells indicating reduced DNA synthesis, consistent with results from cell proliferation assays. In addition, NB4 cells that express low levels of SALL4 have significantly decreased tumorigenecity in immunodeficient mice. Our studies provide a foundation in the development of leukemia stem cell-specific therapy by targeting SALL4. Keywords: ChIP-chip The global targets of SALL4 were determined using a NimbleGen promoter tiling array (2.7kb, information below) in the human cell line NB4. Because SALL4 is thought to play a significant role in leukemogenesis we focused primarily on genes involved in apoptosis and only used the ChIP-chip array as a screening tool to identify potential genes that may bind these genes. Upon identification and verification of apoptosis genes bound by SALL4 we utilized functional assays to determine the effect of SALL4 on these genes. In addition, gene expression profiling was used to determine pathways functionally altered by Sall4 reduction. Source: UCSC Build: HG18 Probe Length: 50-75mer Median Probe Spacing: 100bp Probes per Array: 385,000 Feature Size: 16Î¼m x 16Î¼m Array Dimensions: 17.4mm x 13mm Overall Slide Dimensions: 1" x 3" (25 x 75mm) Recommended Storage: Store arrays desiccated at room temperature. Promoter tiling from 2200bp upstream and 500bp downstream of the transcription start site for RefSeq genes.

Dataset Information

RNA-Seq of mouse embryonic stem cells after knockout and re-introduction of Sall4a

Publications

SALL4 controls cell fate in response to DNA base composition.

Similar Datasets

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