The down regulation of target genes by photo activated DNA nanoscissors.
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 12bp 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.
Tsai
Tsung-Lin
TL
Institute of Basic Medical Sciences, National Cheng Kung University, Tainan 70101, Taiwan.
Shieh
Dar-Bin
DB
Yeh
Chen-Sheng
CS
Tzeng
Yonhua
Y
Htet
Khant
K
Chuang
Kao-Shu
KS
Hwu
Jih Ru
JR
Su
Wu-Chou
WC
D004247:DNA / Q000235:genetics* / Q000378:metabolism*; D053837:DNA Cleavage; D015536:Down-Regulation; D005796:Genes; D006046:Gold / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D006835:Hydrazones / Q000737:chemistry / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry*; D055668:Photochemical Processes
D006835:Hydrazones; D006046:Gold; D004247:DNA