Abstract
Efficient transcriptional programming promises to open new frontiers in regenerative medicine. However, mechanisms by which programming factors transform cell fate are unknown, preventing more rational selection of factors to generate desirable cell types. Three transcription factors, Ngn2, Isl1 and Lhx3, were sufficient to program rapidly and efficiently spinal motor neuron identity when expressed in differentiating mouse embryonic stem cells. Replacement of Lhx3 by Phox2a led to specification of cranial, rather than spinal, motor neurons. Chromatin immunoprecipitation-sequencing analysis of Isl1, Lhx3 and Phox2a binding sites revealed that the two cell fates were programmed by the recruitment of Isl1-Lhx3 and Isl1-Phox2a complexes to distinct genomic locations characterized by a unique grammar of homeodomain binding motifs. Our findings suggest that synergistic interactions among transcription factors determine the specificity of their recruitment to cell type-specific binding sites and illustrate how a single transcription factor can be repurposed to program different cell types.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Basic Helix-Loop-Helix Transcription Factors / metabolism
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Cell Differentiation / physiology*
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Cells, Cultured
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism
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Embryo, Mammalian
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Embryonic Stem Cells
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Gene Expression
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Gene Expression Profiling
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Homeodomain Proteins / metabolism
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Ki-67 Antigen / metabolism
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LIM-Homeodomain Proteins / metabolism
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Mice
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Motor Neurons / cytology
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Motor Neurons / physiology*
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Nerve Tissue Proteins / metabolism
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Oligodendrocyte Transcription Factor 2
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Protein Binding / drug effects
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Protein Binding / genetics
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Protein Structure, Tertiary
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Spinal Cord / cytology
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Stem Cells / cytology
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Stem Cells / drug effects
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Stem Cells / physiology*
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Time Factors
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Transcription Factors / genetics
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Transcription Factors / metabolism*
Substances
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Basic Helix-Loop-Helix Transcription Factors
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DNA-Binding Proteins
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Homeodomain Proteins
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Ki-67 Antigen
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LIM-Homeodomain Proteins
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Lhx3 protein
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Nerve Tissue Proteins
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Neurog2 protein, mouse
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Olig2 protein, mouse
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Oligodendrocyte Transcription Factor 2
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Phox2a protein, mouse
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Transcription Factors
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insulin gene enhancer binding protein Isl-1