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AOC Techniques Continue to Advance: Solving Nuclear Imaging with Left-Handed DNA

Key takeaways

  • Left-handed DNA (L-DNA) is a mirror image of natural right-handed cellular DNA, so it cannot hybridize with genomic DNA and avoids unwanted background in DNA-PAINT imaging.
  • Traditional DNA-PAINT suffers nuclear interference because cellular DNA hybridizes with the imaging probes; swapping in L-DNA oligos substantially reduces that background while keeping the same specificity and multiplexing.
  • Unterauer et al. achieved rapid 13-plex 3D imaging at 15 nm spatial resolution across a 200 × 200 µm² field of view using oligo-conjugated secondary nanobodies.
  • Wider availability of directly conjugated AOCs could accelerate adoption of left-handed DNA-PAINT for nuclear and multiplexed imaging.

The field of Antibody-Oligonucleotide Conjugate (AOC) techniques continues developing at an impressive pace. Following up on our earlier summary of AOC techniques (Scientific Techniques Using Antibody-Oligonucleotide Conjugates), the team at AbOliGo wanted to highlight an elegant improvement to the DNA-PAINT super-resolution imaging technique.

The Problem:

Traditional DNA-PAINT faces interference when imaging the nucleus because cellular DNA can hybridize with the imaging probes, creating unwanted background signals.

Left-handed DNA illustration showing mirror-image property

The Solution:

By using mirror image Left-handed-DNA oligos that cannot interact with natural Right-handed DNA, the team achieved the same specificity and multiplexing capabilities as standard DNA-PAINT while substantially reducing nuclear background.

Building on the 2021 mirror image left-handed DNA-PAINT work by Geertsema et al, Unterauer et al. have just published a streamlined approach combining speed-optimized DNA-PAINT sequences with the left-handed DNA.

Key Achievements:

  • Rapid and efficient 13-plex imaging in 3D with a 15 nm spatial resolution across a 200 × 200 µm² field of view.
  • Used Oligo conjugated (secondary) nanobodies for a lot this work. Wide availability of directly conjugated AOCs could help adoption?

Question

Where else could the Left-handed-DNA trick be applied?

Reference