RNA-Cleaving DNA Thresholder Controlled by Concentrations of miRNA Cancer Marker

Area of Science:

• Molecular Biology
• Biotechnology
• Synthetic Biology

Background:

• Oligonucleotide gene therapy (OGT) agents reduce gene expression by targeting specific mRNAs.
• Low selectivity of OGT agents is a challenge in cancer treatment, as cancer RNA markers are also present in healthy cells, albeit at lower levels.

Purpose of the Study:

• To design a DNA-based construct, the DNA thresholder (DTh), capable of distinguishing cancer cells from healthy cells based on RNA marker concentration.
• To develop a molecular device for selective gene silencing in cancer cells, aiming to trigger cancer cell death.

Main Methods:

• Design of a DNA-based construct (DTh) engineered for RNA-cleaving activity.
• Tuning the DTh's RNA cleavage activity based on the concentration of cancer marker RNA.
• Demonstration of selective RNA cleavage at high cancer marker RNA concentrations and reduced activity at low concentrations.

Main Results:

• The DTh selectively cleaves targeted RNA only at high concentrations of cancer marker RNA.
• The RNA-cleaving activity of DTh can be precisely adjusted by redesigning the construct.
• DTh recognizes cancer marker RNA and cleaves targeted RNA, enabling selective gene suppression in cancer cells.

Conclusions:

• The DNA thresholder (DTh) is a novel computation-inspired molecular device for controlling gene expression.
• DTh demonstrates potential for selective cancer treatment by triggering cancer cell death through targeted gene suppression.
• This technology offers a promising strategy to overcome the selectivity issues in oligonucleotide gene therapy.