Journal
BIOMATERIALS
Volume 31, Issue 25, Pages 6545-6554Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.biomaterials.2010.04.058
Keywords
Gold nanoparticles; Triplex-forming oligonucleotides (TFOs); Hydrazone; DNA cleavge; Gene manipulation
Funding
- National Science Council of Taiwan [NSC 98-3114-M-006-002, DOH99-TD-B-111-002, DOH99-TD-C-111-003, R106]
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An artificial, targeted, light-activated nanoscissor (ATLANS) was developed for precision photonic cleavage of DNA at selectable target sequences. The ATLANS is comprised of nanoparticle core and a monolayer of hydrazone-modified triplex-forming oligonucleotides (TFOs), which recognize and capture the targeted DNA duplex. Upon photo-illumination (lambda = 460 nm), the attached hydrazone scissor specifically cleaves the targeted DNA at a pre-designed nucleotide pair. Electrophoretic mobility shift and co-precipitation assays revealed sequence-specific binding with the short-fragment and long-form plasmid DNA of both TFO and TFO-nanoparticle probes. Upon photo-illumination, ATLANS introduced a precise double-stranded break 12 bp downstream the TFO binding sequence and down-regulated the target gene in HeLa cell system. Gold nanoparticles multiplexed the cutting efficiency and potential for simultaneous manipulation of multiple targets, as well as protected DNA from non-specific photo-damage. This photon-mediated DNA manipulation technology will facilitate high spatial and temporal precision in simultaneous silencing at the genome level, and advanced simultaneous manipulation of multiple targeted genes. (C) 2010 Elsevier Ltd. All rights reserved.
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