Nanocircles

Nanocircles are small circular single-stranded DNA that can be transcribed by phage and bacterial RNA polymerases. These plasmid-like structures were originally developed by Eric T. Kool's lab. The new technology uses a method called rolling circle transcription (RCT) to encode hammerhead, hairpin and hepatitis delta ribozymes.

Rolling Circle Animation Click on Rolling Circles & Artificial Telomeres

Goals

• synthesize efficient self-processing ribozymes
• use ribozymes to regulate genes
• change ribozymes while retaining randomized domain (universality)
• interchange genes for utility
• reinforce importance of secondary structure in cleaving properties

Experimental Design

Figure 1. Structrure of single-stranded DNA nanocircle composed of 63 nucleotides encoding a hammerhead ribozyme and 41 nucleotides of randomized sequences

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Figure 2. Schematic of artificial ribozymes using error prone reverse transcripase PCR

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Results

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http://www.pnas.org/content/vol99/issue1/images/medium/pq0125890004.gif http://www.pnas.org/content/vol99/issue1/images/medium/pq0125890006.gif

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Applications of Ribozymes in Synthetic Systems - Danielle Jordan