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Identification of Key Factors Regulating Self-renewal and Differentiation in EML Hematopoietic Precursor Cells by RNA-sequencing Analysis
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Reprogramming Gene Expression by Targeting RNA-Based Interactions: A Novel Pipeline Utilizing RNA Array Technology.

Charlotte A Henderson1, Helen A Vincent1, Anastasia J Callaghan1

  • 1School of Biological Sciences and Institute of Biological and Biomedical Sciences, University of Portsmouth, Portsmouth, PO1 2DY, United Kingdom.

ACS Synthetic Biology
|July 20, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces a new pipeline to create antisense oligonucleotides (ASOs) that target bacterial small RNAs (sRNAs) and modify gene expression for synthetic biology and antibacterial applications.

Keywords:
RNA arrayRNA−RNA interactionsantisense oligonucleotide (ASO)peptide nucleic acid (PNA)reprogramming gene expressionsmall regulatory RNA (sRNA)

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Area of Science:

  • Molecular Biology
  • Synthetic Biology
  • Antimicrobial Research

Background:

  • Bacterial trans-encoded small RNAs (sRNAs) regulate gene expression via mRNA interactions, crucial for bacterial processes.
  • Modulating these sRNA-mRNA interactions offers potential for synthetic biology and novel antibacterial strategies.

Purpose of the Study:

  • To develop a novel pipeline for designing and validating antisense oligonucleotides (ASOs) targeting bacterial sRNA-mRNA interactions.
  • To reprogram gene expression for synthetic biology and antibacterial applications by modulating RNA-based regulatory networks.

Main Methods:

  • Rational design of anti-sRNA ASOs, synthesized as peptide nucleic acids (PNAs).
  • Development of an RNA array-based assay for in vitro screening of ASO activity.
  • Utilized an Escherichia coli-based reporter assay to validate ASO function in a cellular context.

Main Results:

  • Successfully designed and synthesized anti-sRNA ASOs targeting key sRNA-mRNA interactions.
  • Validated the ability of ASOs to inhibit sRNA-mRNA interactions in vitro.
  • Demonstrated cellular regulatory activity of ASOs in modulating gene expression.

Conclusions:

  • The developed pipeline effectively generates ASOs capable of modulating bacterial gene expression by targeting sRNA-mRNA interactions.
  • This approach holds significant potential for advancing synthetic biology tools and developing new antibacterial therapeutics.
  • The pipeline is anticipated to be broadly applicable for targeting RNA-based regulatory networks across various biological fields.