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Team:Rutgers/main.html
From 2014.igem.org
The Next Step in DNA Synthesis
We believe de novo DNA Synthesis is missing out on a brilliant tool invented by biology that makes stringing nucleotides together extremely quickly and extremely efficiently. We currently use organic solvents and reactive amidites to achieve synthesis, but it may be prudent to begin investigating natural, aqueous environments for synthesis using enzymes evolved alongside nucleic acids for billions of years.
WHYWe've been able to custom-synthesize DNA for over half a century, but our methods have remained relatively unchanged during that timespan. Our current efficiencies are just low enough to limit synthesis to about 150 bases at a time, which brings the cost of DNA for a typical small enzyme above $300 on average [citation needed]. Large protein constructs (like ribosomes) cost several thousands of dollars to code for. The importance of coding price lies in the analogy of Synthetic Biology with Computers - imagine, if building DNA was as cheap as typing up a program, how easy building applications could become? The cost is hugely limiting. We believe the driving factor for cost is the efficiency of reaction. Use biological parts to achieve de novo DNA Synthesis, and there's a chance costs could become negligible. |
HOWTerminal Transferase is a template-independent polymerase that could conceivably eliminate all the complexity of today's synthetic methods. The principles would remain the same - simply add a solution of blocked nucleotides (whichever A, C, G, or T comes next in the sequence) to the immobilized DNA strands, then deblock the terminal nucleotides to allow for the addition of the next nucleotide in the sequence. The rest of the reaction (solvent, pH, phosphate protecting groups) would all become far more similar to physiological conditions, which would eliminate many of the inefficiencies that plague DNA Synthesis today. |
Students
Kenny Kostenbader Chem Eng |
Scott Lazaro Cell Bio & Neurosci |
Wilson Wong Mol Bio |
Jay Patel Chem Eng |
Diego Barreto Biotechnology |
Wesley Okwemba Chem Eng |
Neil Patel Computer Eng |
Harsh Patel Cell Bio & Neurosci |
Gabriel Schimit Chem Eng |
Samantha Ashley Cell Bio & Neurosci |
Faculty
Sagar Khare P.I. |
Andrew Laudisi Lab Manager |
At the NEGEM Meetup (June 20)
(click to see full resolution)
Acknowledgements
Trilink Biotech - Acetylated Nucleotides for free
Dr Jones - synthesis & characterization (Ac2O + LC/MS)
Dr Williams - synthesis & characterization (AcCl + NMR)
Arun Nayar - lab training
BU's NEGEM Meetup - presented and learned a lot from others (see pictures below)
