DNA in Nanochannels - Theory and Applications
Reviewartikel, 2022

Nanofluidic structures have over the last two decades emerged as a powerful platform for detailed analysis of DNA on the kilobase pair length scale. When DNA is confined to a nanochannel, the combination of excluded volume and DNA stiffness leads to the DNA being stretched to near its full contour length. Importantly, this stretching takes place at equilibrium, without any chemical modifications to the DNA. As a result, any DNA can be analyzed, such as DNA extracted from cells or circular DNA, and it is relatively easy to study reactions on the ends of linear DNA. In this comprehensive review, we first give a thorough description of the current understanding of the polymer physics of DNA and how that leads to stretching in nanochannels. We then describe how the versatility of nanofabrication can be used to design devices specifically tailored for the problem at hand, either by controlling the degree of confinement or enabling facile exchange of reagents to measure DNA-protein reaction kinetics. The remainder of the review focuses on two important applications of confining DNA in nanochannels. The first is optical DNA mapping, which provides kilobase pair resolution of the genomic sequence of intact DNA molecules in excess of 100 kilobase pairs in size through labeling strategies that are suitable for fluorescence microscopy. In this section, we highlight solutions to the technical aspects of genomic mapping, rather than recent applications in human genetics, including the use of enzyme-based labeling and affinity-based labeling to produce the genomic maps. The second is DNA-protein interactions, and several recent examples of such studies on DNA compaction, filamentous protein complexes, and reactions with the chain ends are presented. Taken together, these two applications demonstrate the power of DNA confinement and nanofluidics in genomics, molecular biology and biophysics.

nanofluidics

Optical DNA mapping

DNA

single molecules

polymer physics

Författare

Karolin Frykholm

Chalmers, Biologi och bioteknik, Kemisk biologi

Vilhelm Müller

Chalmers, Biologi och bioteknik, Kemisk biologi

Sriram Kesarimangalam

Chalmers, Biologi och bioteknik, Kemisk biologi

Kevin D. Dorfman

University of Minnesota

Fredrik Westerlund

Chalmers, Biologi och bioteknik, Kemisk biologi

Quarterly Reviews of Biophysics

0033-5835 (ISSN) 1469-8994 (eISSN)

Vol. 55 e12

Ämneskategorier

Biokemi och molekylärbiologi

Biofysik

Annan industriell bioteknik

DOI

10.1017/S0033583522000117

PubMed

36203227

Mer information

Senast uppdaterat

2024-03-07