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A Multi-step Transcriptional and Chromatin State Cascade Underlies Motor Neuron Programming from Embryonic Stem Cells.

Cell stem cell | 2017

Direct cell programming via overexpression of transcription factors (TFs) aims to control cell fate with the degree of precision needed for clinical applications. However, the regulatory steps involved in successful terminal cell fate programming remain obscure. We have investigated the underlying mechanisms by looking at gene expression, chromatin states, and TF binding during the uniquely efficient Ngn2, Isl1, and Lhx3 motor neuron programming pathway. Our analysis reveals a highly dynamic process in which Ngn2 and the Isl1/Lhx3 pair initially engage distinct regulatory regions. Subsequently, Isl1/Lhx3 binding shifts from one set of targets to another, controlling regulatory region activity and gene expression as cell differentiation progresses. Binding of Isl1/Lhx3 to later motor neuron enhancers depends on the Ebf and Onecut TFs, which are induced by Ngn2 during the programming process. Thus, motor neuron programming is the product of two initially independent transcriptional modules that converge with a feedforward transcriptional logic.

Pubmed ID: 27939218 RIS Download

Associated grants

  • Agency: NHGRI NIH HHS, United States
    Id: DP2 HG009623
  • Agency: NICHD NIH HHS, United States
    Id: R01 HD079682

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This is a list of tools and resources that we have found mentioned in this publication.


Enrichr (tool)

RRID:SCR_001575

A web-based gene list enrichment analysis tool that provides various types of visualization summaries of collective functions of gene lists. It includes new gene-set libraries, an alternative approach to rank enriched terms, and various interactive visualization approaches to display enrichment results using the JavaScript library, Data Driven Documents (D3). The software can also be embedded into any tool that performs gene list analysis. System-wide profiling of genes and proteins in mammalian cells produce lists of differentially expressed genes / proteins that need to be further analyzed for their collective functions in order to extract new knowledge. Once unbiased lists of genes or proteins are generated from such experiments, these lists are used as input for computing enrichment with existing lists created from prior knowledge organized into gene-set libraries.

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PANTHER (tool)

RRID:SCR_004869

System that classifies genes by their functions, using published scientific experimental evidence and evolutionary relationships to predict function even in absence of direct experimental evidence. Orthologs view is curated orthology relationships between genes for human, mouse, rat, fish, worm, and fly.

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UCSC Genome Browser (tool)

RRID:SCR_005780

Portal to interactively visualize genomic data. Provides reference sequences and working draft assemblies for collection of genomes and access to ENCODE and Neanderthal projects. Includes collection of vertebrate and model organism assemblies and annotations, along with suite of tools for viewing, analyzing and downloading data.

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LIMMA (tool)

RRID:SCR_010943

Software package for the analysis of gene expression microarray data, especially the use of linear models for analyzing designed experiments and the assessment of differential expression.

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SAM format (tool)

RRID:SCR_012093

A generic alignment format for storing read alignments against reference sequences, supporting short and long reads (up to 128 Mbp) produced by different sequencing platforms.

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Bowtie 2 (tool)

RRID:SCR_016368

Ultrafast and memory efficient tool for aligning sequencing reads to long reference sequences. Supports gapped, local, and paired end alignment modes. More suited to finding longer, gapped alignments in comparison with original Bowtie method.

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PANTHER (tool)

RRID:SCR_015893

System that classifies genes by their functions, using published scientific experimental evidence and evolutionary relationships to predict function even in absence of direct experimental evidence. Orthologs view is curated orthology relationships between genes for human, mouse, rat, fish, worm, and fly.

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EBF2 antibody (antibody)

RRID:AB_2097415

This polyclonal targets EBF2 antibody

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H3K27ac-human (antibody)

RRID:AB_2118291

This polyclonal targets H3K27ac

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H3K4me3-dmelanogaster (antibody)

RRID:AB_2615077

This unknown targets H3K4me3

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H3K4me2-human (antibody)

RRID:AB_2560996

This polyclonal targets H3K4me2

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Neurogenin 2 (C-16) (antibody)

RRID:AB_2149513

This polyclonal targets NEUROG2

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V5 tag antibody - ChIP Grade (antibody)

RRID:AB_443253

This polyclonal targets V5 tag antibody - ChIP Grade

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Human ONECUT2/OC-2 Affinity Purified Polyclonal Ab (antibody)

RRID:AB_10640365

This polyclonal targets Human ONECUT2/OC-2 Affinity Purified Ab

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Human/Mouse EBF-2 Affinity Purified Polyclonal Ab (antibody)

RRID:AB_10972102

This unknown targets Human/Mouse EBF-2 Affinity Purified Ab

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Anti-beta-Tubulin III antibody produced in rabbit (antibody)

RRID:AB_262133

This polyclonal targets beta-Tubulin III antibody produced in rabbit

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Ki67 antibody - Proliferation Marker (antibody)

RRID:AB_443209

This polyclonal targets Ki67 antibody - Proliferation Marker

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Oct3/4 Antibody (C-10) (antibody)

RRID:AB_628051

This monoclonal targets Oct-3/4

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Bowtie (software resource)

RRID:SCR_005476

Software ultrafast memory efficient tool for aligning sequencing reads. Bowtie is short read aligner.

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RSEM (software resource)

RRID:SCR_013027

Software package for quantifying gene and isoform abundances from single end or paired end RNA Seq data. Accurate transcript quantification from RNA Seq data with or without reference genome. Used for accurate quantification of gene and isoform expression from RNA-Seq data.

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GREAT: Genomic Regions Enrichment of Annotations Tool (software resource)

RRID:SCR_005807

Data analysis service that predicts functions of cis-regulatory regions identified by localized measurements of DNA binding events across an entire genome. Whereas previous methods took into account only binding proximal to genes, GREAT is able to properly incorporate distal binding sites and control for false positives using a binomial test over the input genomic regions. GREAT incorporates annotations from 20 ontologies and is available as a web application. The utility of GREAT extends to data generated for transcription-associated factors, open chromatin, localized epigenomic markers and similar functional data sets, and comparative genomics sets. Platform: Online tool

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BEDOPS (software resource)

RRID:SCR_012865

A suite of tools to address common questions raised in genomic studies - mostly with regard to overlap and proximity relationships between data sets.

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Flexbar (software resource)

RRID:SCR_013001

Flexible barcode and adapter removal for sequencing platforms.

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BEDTools (software resource)

RRID:SCR_006646

A powerful toolset for genome arithmetic allowing one to address common genomics tasks such as finding feature overlaps and computing coverage. Bedtools allows one to intersect, merge, count, complement, and shuffle genomic intervals from multiple files in widely-used genomic file formats such as BAM, BED, GFF/GTF, VCF. While each individual tool is designed to do a relatively simple task (e.g., intersect two interval files), quite sophisticated analyses can be conducted by combining multiple bedtools operations on the UNIX command line.

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SAMTOOLS (software resource)

RRID:SCR_002105

Original SAMTOOLS package has been split into three separate repositories including Samtools, BCFtools and HTSlib. Samtools for manipulating next generation sequencing data used for reading, writing, editing, indexing,viewing nucleotide alignments in SAM,BAM,CRAM format. BCFtools used for reading, writing BCF2,VCF, gVCF files and calling, filtering, summarising SNP and short indel sequence variants. HTSlib used for reading, writing high throughput sequencing data.

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