Single Molecule Localization and Discrimination of DNA–Protein Complexes by Controlled Translocation Through Nanocapillaries
Through the use of optical tweezers we performed controlled translocations of DNA-protein complexes through nanocapillaries. We used RNA polymerase (RNAP) with two binding sites on a 7.2 kbp DNA fragment and a dCas9 protein tailored to have five binding sites on lambda-DNA (48.5 kbp). Measured localization of binding sites showed a shift from the expected positions on the DNA that we explained using both analytical fitting and a stochastic model. From the measured force versus stage curves we extracted the non equilibrium work done during the translocation of a DNA-protein complex and used it to obtain an estimate of the effective charge of the complex. In combination with conductivity measurements, we provided a proof of concept for discrimination between different DNA protein complexes simultaneous to the localization of their binding sites.
Cite this item
Bulushev, R. D., Marion, S., Petrova, E., Davis, S. J., Maerkl, S. J., & Radenovic, A. (2016). Single Molecule Localization and Discrimination of DNA–Protein Complexes by Controlled Translocation Through Nanocapillaries. Nano Letters, 16(12), 7882–7890. https://doi.org/10.1021/acs.nanolett.6b04165
Click Copy to copy the citation, or select the text manually.
Nano_letters_Bulushev_2016_SI.pdf
openaccess
1.06 MB
Adobe PDF
3534f13b011ab3b6b9f3340eeb4390c0
Nano_letters_Bulushev_2016_ms.pdf
openaccess
2.8 MB
Adobe PDF
56b66506632d3de730b42fbd0bdc93e4