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
http://www.pnas.org/content/vol99/issue1/images/medium/pq0125890001.gif
Figure 2. Schematic of artificial ribozymes using error prone reverse transcripase PCR
http://www.pnas.org/content/vol0/issue2001/images/data/012589099/DC1/5890Fig9.gif
Results
http://www.pnas.org/content/vol99/issue1/images/medium/pq0125890002.gif
http://www.pnas.org/content/vol99/issue1/images/medium/pq0125890004.gif http://www.pnas.org/content/vol99/issue1/images/medium/pq0125890006.gif
http://www.pnas.org/content/vol99/issue1/images/medium/pq0125890007.gif
http://www.pnas.org/content/vol99/issue1/images/medium/pq0125890008.gif
Applications of Ribozymes in Synthetic Systems - Danielle Jordan