Single-molecule optical-tweezers

Using single-molecule optical-tweezers we catch and manipulate individual molecules of DNA. We use integrated confocal microscopy to directly image DNA stained with fluorescent dyes or fluorescently labelled proteins interacting with DNA.

Lambda-phage DNA labelled with Sytox Orange

CRISPR/Cas9 binding at off-target sites when DNA is stretched (Newton et al, 2019)

Watching individual proteins in action

Using optical-tweezers we can directly observe individual proteins interacting with a single piece of DNA in real-time. This allows us to address important questions about the mechanism of DNA-protein interactions.

For example we are able to directly observe binding of the gene-editing protein CRISPR/Cas9, revealing that changes in DNA structure can dramatically increase off-target binding.

Different DNA substrates

As well as using double-stranded DNA we are able to generate a wide variety of different DNA substrates to study specific biological questions. We can use single-stranded DNA and DNA containing stranded gaps, DNA with complex structures such as Holliday junctions, RNA hairpins, and DNA containing different types of DNA damage.

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