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Charles Olea, PhD

Research Associate
Sept 2010 – present
PhD, 2010, UC Berkeley
Research: Autocatalytic aptazymes for detection of biomarkers
E-mail: colea@scripps.edu

All biopolymers in living organisms exist in one enantiomeric form. The genetic material is composed of D-nucleotides and the protein machinery that brings about replication of the genetic material is composed of L-amino acids. Therefore replication, which is the fundamental process of life, is itself homochiral.

Recently an RNA-based synthetic genetic system was developed that can undergo self-sustained replication. This system involves RNA enzymes that produce additional copies of themselves by joining oligonucleotide building blocks. The replicating RNA molecules carry genetic information and amplify exponentially without the aid of proteins.

Olea Figure

The self-replicating RNA enzymes have been engineered to function as “aptazymes”, which undergo exponential amplification in a ligand-dependent manner. These autocatalytic aptazymes have potential applications in molecular diagnostics, analogous to quantitative PCR but for a broader range of targets. However, their use had been limited due to their susceptibility to degradation by ribonucleases, which are present in all biological samples. To address this limitation, the autocatalytic aptazymes have been constructed entirely of L-ribonucleotides. As expected, the L-RNA molecules have the same physical and chemical properties as their D counterparts, but are completely resistant to ribonucleases. Autocatalytic L-aptazymes are currently being developed to detect various peptide and protein disease biomarkers.

Methods are in development for the real-time fluorescence measurement of exponential RNA amplification. One such method relies on a combination of fluorophores and quenchers to report the yield of the amplified products. The availability of multiple fluorophores that can be coupled to individual biomarker-specific replicators should provide a multiplexed approach for real-time molecular diagnostics.

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