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Epigenetics is the study of inherited changes in a cell's phenotype without changing the DNA sequences. It provides a form of memory for the differential gene expression pattern to maintain cell lineage, position-effect variegation, dosage compensation, and maintenance of chromatin structures such as telomeres and centromeres. For example, the structure and location of the centromere on chromosomes are epigenetically inherited. Its functionality is not dictated or ensured by the underlying...
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Investigation of the Transcriptional Role of a RUNX1 Intronic Silencer by CRISPR/Cas9 Ribonucleoprotein in Acute Myeloid Leukemia Cells
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Cis-regulatory elements are harbored in Intron5 of the RUNX1 gene.

Boris Rebolledo-Jaramillo, Ricardo A Alarcon, Valentina I Fernandez

  • 1Departamento de Bioquimica y Biologia Molecular, Facultad de Ciencias Biologicas, Universidad de Concepcion, Concepcion, Chile. sgutierr@udec.cl.

BMC Genomics
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Summary

RUNX1 gene intron 5 contains regulatory elements crucial for acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL). These conserved non-coding sequences show cell-type specific transcriptional activity, suggesting a role in gene regulation.

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Area of Science:

  • Genetics
  • Molecular Biology
  • Cancer Research

Background:

  • The RUNX1 gene is frequently altered in acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL) via chromosomal translocations.
  • The t(8;21) translocation, common in AML, involves breakpoints in RUNX1 intron 5 and ETO intron 1.
  • DNase I hypersensitive sites (DHS) in these regions suggest the presence of regulatory elements.

Purpose of the Study:

  • To investigate potential regulatory elements within intron 5 of the human RUNX1 gene.
  • To identify and characterize conserved non-coding sequences in RUNX1 intron 5.

Main Methods:

  • Comparative genomics to identify conserved non-coding sequences.
  • Cloning of candidate regions into a reporter plasmid (pGL-3).
  • Transfection assays to evaluate transcriptional regulatory activity in a cell-type specific manner.

Main Results:

  • Nine evolutionarily conserved non-coding sequences were identified in RUNX1 intron 5.
  • Two of these regions demonstrated the ability to regulate reporter gene transcription.
  • The observed transcriptional regulation was independent of distance and position, and exhibited cell-type specificity.

Conclusions:

  • RUNX1 intron 5 harbors multiple conserved non-coding sequences.
  • These sequences possess cis-regulatory potential, capable of modulating transcriptional activity.
  • These findings contribute to understanding the regulatory landscape of the RUNX1 gene in leukemia.