Journal
NATURE PROTOCOLS
Volume 10, Issue 10, Pages 1508-1524Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2015.078
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Funding
- National Key Scientific Program of China [2011CB911000]
- National Natural Science Foundation of China (NSFC) [21325520, 21327009, 21221003, J1210040, 21177036, 21135001]
- China National Instrumentation Program [2011YQ03012412]
- US National Institutes of Health [GM079359, CA133086]
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We describe a comprehensive protocol for the preparation of multifunctional DNANA nanostructures termed nanoflowers (NFs), which are self-assembled from long DNA building blocks generated via rolling-circle replication (RCR) of a designed template. NF assembly is driven by liquid crystallization and dense packaging of building blocks, which eliminates the need for conventional Watson-Crick base pairing. As a result of dense DNA packaging, NFs are resistant to nuclease degradation, denaturation or dissociation at extremely low concentrations. By manually changing the template sequence, many different functional moieties including aptamers, bioimaging agents and drug-loading sites could be easily integrated into NF particles, making NFs ideal candidates for a variety of applications in biomedicine. In this protocol, the preparation of multifunctional DNA NFs with highly tunable sizes is described for applications in cell targeting, intracellular imaging and drug delivery. Preparation and characterization of functional DNA NFs takes similar to 5 d; the following biomedical applications take similar to 10 d.
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