Biomedical subjects

Joanne Macdonald

Publications and source records attributed to Joanne Macdonald.

5 recordsLinked to original sources

PubMed2006

Deoxyribozyme-based three-input logic gates and construction of a molecular full adder.

We have developed an array of seven deoxyribozyme-based molecular logic gates that behaves as a full adder in a single solution, with three oligonucleotides as inputs and two independent fluorogenic cleavage reactions as carry and sum outputs. The sum output consisted of four new deoxyribozyme-based logic gates: an ANDAND gate and three ANDNOTANDNOT gates. These gates required the design of a generic three-input deoxyribozyme-based logic gate that can use any three-way combination of activating or inactivating inputs. This generic gate design utilizes an additional inverting element that hybridizes to convert YES logic into NOT logic and vice versa. The system represents the first solution-phase, single test tube, enzymatic full adder and shows the complexity of control over molecular scale events that can be achieved with deoxyribozyme-based logic gates. Similar systems could be applied to control autonomous therapeutic and diagnostic devices.

Base Sequence↗

PubMed2006

Medium scale integration of molecular logic gates in an automaton.

The assembly of molecular automata that perform increasingly complex tasks, such as game playing, presents an unbiased test of molecular computation. We now report a second-generation deoxyribozyme-based automaton, MAYA-II, which plays a complete game of tic-tac-toe according to a perfect strategy. In silicon terminology, MAYA-II represents the first "medium-scale integrated molecular circuit", integrating 128 deoxyribozyme-based logic gates, 32 input DNA molecules, and 8 two-channel fluorescent outputs across 8 wells.

Algorithms↗

PubMed2006

Solution-phase molecular-scale computation with deoxyribozyme-based logic gates and fluorescent readouts.

Recent development of solution-phase molecular-scale Boolean calculations using deoxyribozymes is potentially an important step toward the development of autonomous therapeutic and diagnostic devices. Here, the construction of basic YES, AND, ANDNOT, and ANDANDNOT deoxyribozyme-based logic gates is described. Protocols for testing gate activity using fluorescent oligonucleotide probes have been provided, and pointers for gate optimization are included.

Base Sequence↗

PubMed2005

Deoxyribozyme-based ligase logic gates and their initial circuits.

A complete set (YES, NOT, AND, and ANDNOT) of molecular scale logic gates based on ligase deoxyribozymes was constructed. The activity of these gates was visualized through the formation of cascades with downstream phosphodieseterase YES gates, which performed fluorogenic cleavage.

Base Sequence↗

PubMed2005

NS1 protein secretion during the acute phase of West Nile virus infection.

The West Nile virus (WNV) nonstructural protein NS1 is a protein of unknown function that is found within, associated with, and secreted from infected cells. We systematically investigated the kinetics of NS1 secretion in vitro and in vivo to determine the potential use of this protein as a diagnostic marker and to analyze NS1 secretion in relation to the infection cycle. A sensitive antigen capture enzyme-linked immunosorbent assay (ELISA) for detection of WNV NS1 (polyclonal-ACE) was developed, as well as a capture ELISA for the specific detection of NS1 multimers (4G4-ACE). The 4G4-ACE detected native NS1 antigens at high sensitivity, whereas the polyclonal-ACE had a higher specificity for recombinant forms of the protein. Applying these assays we found that only a small fraction of intracellular NS1 is secreted and that secretion of NS1 in tissue culture is delayed compared to the release of virus particles. In experimentally infected hamsters, NS1 was detected in the serum between days 3 and 8 postinfection, peaking on day 5, the day prior to the onset of clinical disease; immunoglobulin M (IgM) antibodies were detected at low levels on day 5 postinfection. Although real-time PCR gave the earliest indication of infection (day 1), the diagnostic performance of the 4G4-ACE was comparable to that of real-time PCR during the time period when NS1 was secreted. Moreover, the 4G4-ACE was found to be superior in performance to both the IgM and plaque assays during this time period, suggesting that NS1 is a viable early diagnostic marker of WNV infection.

Animals↗