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A single-input binary counting module based on serine integrase site-specific recombination.

Jia Zhao¹, Alexandra Pokhilko¹, Oliver Ebenhöh^2,3

¹Institute of Molecular, Cell and Systems Biology, University of Glasgow, Bower Building, Glasgow G12 8QQ, Scotland.

Nucleic Acids Research

|April 9, 2019

Summary

This summary is machine-generated.

Researchers developed a DNA-based molecular counter that reliably records cellular events. This innovative device uses a DNA latch mechanism, enabling precise counting for biological and biotechnological applications.

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Area of Science:

Synthetic Biology
Molecular Biology
Biotechnology

Background:

Cellular event counting is crucial for experimental biology and biotechnology.
Existing methods for tracking cellular events can be limited in precision and scope.

Purpose of the Study:

To engineer a DNA-based molecular device capable of counting discrete events within a single cell.
To develop a switchable DNA latch system that responds to repeated stimuli.

Main Methods:

Utilized ϕC31 integrase and recombination directionality factor (RDF) to invert a DNA segment.
Implemented a feedback loop where RDF expression is controlled by the latch state.
Incorporated a transcriptional repressor delay circuit for precise switching per input pulse.
Employed mathematical modeling for optimizing DNA recombination and gene expression dynamics.

Main Results:

Demonstrated a DNA latch that efficiently switches states upon repeated stimuli, acting as a molecular counter.
Achieved precise inversion of a DNA segment mediated by integrase and RDF.
Validated the functionality of the delay circuit ensuring single inversion per stimulus.
Showcased the potential for chaining multiple latches to create binary counters.

Conclusions:

The developed DNA latch serves as a robust molecular counter for cellular events.
This system offers a novel platform for quantitative biological measurements.
Chaining these latches could lead to complex counting devices like binary ripple counters.