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Related Concept Videos

DNA Isolation01:24

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...

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Long-Lasting and Responsive DNA/Enzyme-Based Programs in Serum-Supplemented Extracellular Media.

Jean-Christophe Galas1, André Estevez-Torres1, Marc Van Der Hofstadt1

  • 1Sorbonne Université, CNRS, Institut de Biologie Paris-Seine (IBPS), Laboratoire Jean Perrin (LJP), F-75005, Paris, France.

ACS Synthetic Biology
|February 8, 2022
PubMed
Summary

DNA molecular programs now function in serum for extended periods. Novel three-letter code networks and buffers overcome nuclease degradation, enabling new in vitro diagnostic tools.

Keywords:
DNA molecular programsendonucleaseliving cellsresponsive networksserum

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Area of Science:

  • Biochemistry
  • Synthetic Biology
  • Molecular Engineering

Background:

  • DNA molecular programs offer versatile biomolecular sensing and actuation for pharmaceuticals.
  • Serum nucleases limit DNA program functionality in vitro, causing rapid network deterioration.

Purpose of the Study:

  • To develop DNA molecular programs that are functional and stable in serum-rich environments.
  • To enhance the biocompatibility and responsiveness of DNA programs for in vitro applications.

Main Methods:

  • Implementation of three-letter code DNA networks to suppress parasitic amplification.
  • Development of a novel buffer to improve biocompatibility and responsiveness.
  • Demonstration of serum-supplemented extracellular DNA programs in the presence of living cells.

Main Results:

  • DNA/enzyme programs remained functional in 10% serum for at least 3 days using three-letter code networks.
  • The new buffer enhanced biocompatibility and maintained responsiveness to molecular changes.
  • Extracellular DNA programs showed rapid responses to molecular inputs (6x faster than cell division) and sustained function over three cell divisions.

Conclusions:

  • Developed robust DNA molecular programs functional in serum for over 3 days, overcoming nuclease degradation.
  • Demonstrated the potential for in situ biomolecular characterization tools in serum-demanding in vitro models.
  • Paved the way for extracellular synthetic biology tools and autonomous in vitro models through chemical reactivity coupling.