Single-molecule detection enables direct characterization of annealing/melting kinetics of nucleic acids without the need for synchronization of molecular states, but the current experiments are not carried out in a native cellular context. Here we describe an integrated 3D single-molecule tracking and lifetime measurement method that can follow individual DNA molecules diffusing inside a mammalian cell and observe multiple annealing and melting events on the same molecules. By comparing the hybridization kinetics of the same DNA strand , we found the association constants can be 13- to 163-fold higher in the molecular crowding cellular environment.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7179573 | PMC |
| http://dx.doi.org/10.1021/jacs.9b08036 | DOI Listing |
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