Synthetic Biology is full of promises ranging from discovery and production of new drugs, targeted therapies modifying organisms such as yeast or E. coli or creating totally new ones, higher crop yields, CO2 sequestration to bio-energy projects. However, these goals have been difficult to reach primarily due to the difficulty of De Novo gene synthesis and synthesis of new control and regulatory metabolic pathways all the way to synthetic chromosomes from chemically synthesized DNA. Just like any reversible chemical reaction DNA fragment synthesis by sequentially adding the monomers (A,C,G,T) gives less than 100% yield at each step( typical stepwise yields are ~97-99%) as a result the longer the length of DNA to be synthesized the smaller is the fraction of pure product at the end. Elimination of the errors take significant time and money. The goal of this project is to provide error free long oligonucleotides (100 to 300 mers) at a cost lower than the impure DNA fragments of today to unlock the potential of synthetic biology. We are proposing to make libraries of clonally amplified and sequence verified long oligonucleotides. Our long term objectives are to implement a commercial service of affordable custom synthesis of sequence-verified long oligonucleotide libraries. During Phase I, we will 1) to demonstrate that single oligonucleotide molecules can be clonally amplified and sequence verified, 2) to demonstrate that beads bearing clonal amplifications can be captured on a microarray bearing sequence-specific probes and 3) to demonstrate that sequence verified oligonucleotides enable gene assembly with ten fold reduced error rate. During Phase II, we will 1) to develop a protocol/device to normalize the number of each oligonucleotide surveyed from a library, 2) to establish a standard operating procedure for the production of large libraries of sequence-verified oligonucleotides and 3) to determine the maximum oligonucleotide length that could be offered as a commercial product.

Public Health Relevance

The unprecedented availability of affordable custom libraries of sequence verified long oligonucleotides will enable faster, easier and cheaper gene and large DNA fragment assembly with major applications in Synthetic Biology. This technology will undoubtedly bolster the discovery of new cellular mechanisms, molecular therapeutic tools, or even drugs, ultimately benefiting the society.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Small Business Innovation Research Grants (SBIR) - Phase II (R44)
Project #
1R44GM105192-01
Application #
8455467
Study Section
Special Emphasis Panel (ZRG1-IMST-G (10))
Program Officer
Fabian, Miles
Project Start
2012-09-24
Project End
2013-09-23
Budget Start
2012-09-24
Budget End
2013-09-23
Support Year
1
Fiscal Year
2012
Total Cost
$307,411
Indirect Cost
Name
Biodiscovery, LLC
Department
Type
DUNS #
611643813
City
Ann Arbor
State
MI
Country
United States
Zip Code
48105