Wednesday 4 November 2020

Storage Wars: A battle of nucleic proportions

Reading and writing digital information in TNA

DNA molecules have become a popular information storage system but are susceptible to degradation in a cell-like environment. In this paper, the authors suggest the use of the unnatural genetic framework based on TNA. The backbone structure of this unnatural genetic polymer is recalcitrant to nuclease digestion. 23 kilobytes of digital information were stored in TNA and recovered with perfect accuracy after exposure to biological nucleases that destroyed equivalent DNA messages.

https://pubs.acs.org/doi/pdf/10.1021/acssynbio.0c00361

Metastable hybridization-based DNA information storage to allow rapid and permanent erasure

The authors argue that data stored in DNA in the traditional manner is hard to wipe clean - it requires enzymes, UV or extreme heat. They propose a stable but easily-erasable memory, in which single strands carry a memory location domain and the actual contents of the memory. Multiple distinct strands with the same memory location domain are included, but only one is prepared with a "truth" flag (a strand that is complementary to the memory location domain) whilst the rest are prepared with "false" flags (chemically modified strands). Data can be read out by performing PCR on the system, since the modified strands that constitute the false flags cannot be extended. Data can be easily wiped through a short heating protocol, which will randomise the flags very quickly. While the method achieves this functionality pretty well, it is limited - there is no scope to erase and then re-write data, for example. 

https://www.nature.com/articles/s41467-020-18842-6

Exponential volume dependence of entropy-current fluctuations at first-order phase transitions in chemical reaction networks

In this paper, the divergence of the fluctuations of a chemical switch as a function of system size is quantified and a critical exponent is calculated. The chemical reaction scheme of Schlögl model is studied. Here, the steady state number of a certain chemical species X displays a first order phase transition as the reaction rates are varied. This paper uses various methods to analyse the chemical master equation (CME) at the transition and calculate the variance to find an exponential dependence on system size with a given exponent. This provides a very readable example of a few methods to analyse CMEs.

https://journals.aps.org/pre/abstract/10.1103/PhysRevE.102.022101

Single-particle cryo-EM at atomic resolution

Cryogenic electron microscopy is used to determine the atomic-scale structure of biomolecules without needing to form a crystal of the sample, as is required for x-ray crystallography. In single-particle cryo-EM, a sample containing a high concentration of the molecule of interest is flash frozen and imaged repeatedly with a beam of single electrons. The scattered electrons, that are captured by a camera, form an image of the sample which is used to reconstruct the shape of the molecule of interest. Here the authors report three improvements that brought the resolution of cryo-EM below 2 Angstroms; an electron source with a narrow energy spread that reduced chromatic aberrations, a more effective energy filter that can separate out useful elastically and useless inelastically scattered electrons, and an advanced camera and reconstruction algorithm capable of capturing scattered electrons with high spatio-temporal resolution. In a reconstruction of a 5-fold symmetric human membrane channel protein, small molecule coordination, alternative amino acid conformations and side chain variations were resolved. Further, using an advanced reconstruction method of a 24-fold symmetric mouse protein, the scattering potential of hydrogen atoms could be resolved in the most ordered parts of the structure, enabling analysis of the hydrogen-bonding network in the protein. Atomic resolution cryo-EM will provide insights into the mechanical function of biomolecules and improve structure-based drug discovery.

https://www.nature.com/articles/s41586-020-2829-0

The evolution of DNA-templated synthesis as a tool for materials discovery

Control of reactivity and product structure is one of the main challenges during the design of chemical reactions. A method to introduce said control is DNA-templated synthesis (DTS), where DNA-bound chemicals are assembled over a complementary DNA strand that acts as a template. The focus of this review is the potential application of DTS for the directed evolution of chemicals, by several cycles of template mutation and selection. The authors enumerate all the possible DTS mechanisms that allow the implementation of selection cycles, the chemical reactions templated with said mechanisms, and the challenges they face.

https://pubs.acs.org/doi/10.1021/acs.accounts.7b00280

Nanopore-based DNA hard drives for rewritable and secure data storage

Solid-state nanopores have been demonstrated to be powerful molecular sensors for detecting nanoscale objects. In this article, a long DNA scaffold is use as a “hard drive” to write, erase, and rewrite data on, and read it out by passing the strand through a solid-state nanopore. The data sites are functionalised with small DNA-overhangs which can be hybridised with unique complementary biotinylated strands. The unhybridised and hybridised strands generate weaker and stronger current signals respectively when passed through the nanopore which can be read as 0 or 1. Thus, a combination of data sites can encode meaningful data in binary format. To erase the data, another fuel strand is added which utilises a short toehold at the end of the biotynilated strands to detach it from the overhangs which can be reused again to encode different data. To refine this approach, the group has also demonstrated the utilization of two separate encoding sites (address sites and data sites) simultaneously to encode both the data and its sequence. Moreover, they showed the use of a physical “key” to encrypt the data on the DNA hard drive without which the readout does not match the actual encoded data.

https://pubs.acs.org/doi/full/10.1021/acs.nanolett.0c00755​​​​​​​

Toehold-mediated strand displacement reaction for dual-signal electrochemical assay of Apolipoprotein E genotyping

Apolipoprotein E (ApoE) is a polymorphic gene which has been identified as an important genetic determinant for Alzheimer's disease. In the human population six genotypes of ApoE exist, with distinct risks associated with each. In this paper, the authors developed a toehold-mediated strand displacement-based approach to genotype ApoE. Using an electrochemical detection system, the authors could detect the presence of absense of mutations at 2 codons within the gene. This system allowed for systematic characterisation of ApoE genotype. 

https://pubs.acs.org/doi/pdf/10.1021/acssensors.0c01511


Proton-driven transformable nanovaccine for cancer immunotherapy

They developed a polymer-peptide conjugate that forms nanoscale sphere at normal pH. The polymer gets uptaken by cancer cells. When the polymer is at the endosome, it changes conformation to micrometre sized sheet and destroys the endosome. Then the peptide gets presented outside of the cell to trigger an inflammation response.


Light-activated signaling in DNA-encoded sender–receiver architectures

In this last work from De Greef's lab, the authors present the last developments on their compartmentalized DNA strand displacement platform BIO-PC. More precisely, they show how with the introduction of a photo-cleaving group in a DNA duplex, combined with different tunable variables, this system allows them to implement localized spatial activation of strand displacement circuits with controlled diffusion.  Based on this, they demonstrate how catalytic activation can transmit a given signal further than conventional activation and that this system allows them to build a spatially-coded AND gate.

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