Team Acknowledgments

Our project was selected from a pool of 10 possible project ideas submitted by both team members and advisers. Team members decided to focus on zinc finger proteins because of the technologies (chip DNA synthesis and MAGE) available to us: we believed that developing new zinc finger proteins was a way to use both technologies to their greatest possible extent. Also, we knew that little to no zinc finger research was being conducted at Harvard, with the exception of Dr. Keith Joung's lab at Harvard Medical School. The original idea of zinc finger proteins was contributed by the Harvard iGEM 2011 advisors and mentors, including but not limited to, Jagesh Shah, Srivatsan Raman, Daniel Goodman, Noah Taylor, Jameson Rogers, Joyce Yang, and Jun Li.

This project was made possible by support from Dr. George Church's lab at the Wyss Institute for Biologically Inspired Engineering as well as Alain Viel at the Harvard Department of Molecular and Cellular Biology. Without their valuable guidance and great patience, we never would have been able to achieve what have done so far. While our teaching fellows - members of the Church Lab - gave us many opportunities, it also made sure that we achieved our goals and results entirely via our own work: our teaching fellows provided inspiration, team members achieved what we did through our own perspiration.

Although the project is the first to combine several key technologies in novel ways, these technologies were developed outside of Harvard iGEM. The multiplex automated genome engineering (MAGE) method was developed by Harris Wang et al[2]. Chip-based DNA synthesis method was developed by Sri Kosuri et al[1], and the actual oligo synthesis was generously provided by Agilent Technologies, a sponsor of iGEM. Lambda red was originally developed by Yu et al[3]. We would like to make it clear that we do not claim any credit whatsoever for the creation of the technologies used in this project, just as we claim no credit for the invention of PCR or other technologies used. Just as each iGEM team has utilized a variety of synthetic biology tools in conducting research, for us these technologies are tools we have harnessed for a purpose beyond their mere use. Our project and Foundational Advance proposal rest on our novel integration and application of these multiple technologies in concert, and subsequent application of this system to generate novel zinc finger proteins as a proof of concept. Additionally, it should be noted that the developers of these technologies acted purely as consultants and sounding boards for questions on use and troubleshooting: all of the

We owe great thanks to Alain Viel for his flawless management of the logistics of iGEM, including but not limited to providing lab space, ordering lab supplies when we needed them, arranging our trip to the Americas Regional and World Jamborees, and for his excellent taste in footwear. Furthermore, we would also like to thank our lab manager Andrew Cumming for all of his practical lab advice and for helping to make everything run smoothly in the day to day logistics of lab.

Jagesh Shah provided us with excellent advice on both our project and the iGEM competition as a whole: what to expect, how to improve our presentation, and not to take ourselves too seriously. We also owe Jagesh's children Harvard iGEM t-shirts for depriving them of Jagesh's company when several of our meetings ran late into the night.

Team Member Acknowledgments

All project work was done by team members, divided according to their skills and expertise, with primary areas of work including:

Kristin Barclay
Bioinformatics, chip sequences design and programming, majority of wiki coding and design, human practices

Justin Chew
Bioinformatics, ZF target selection, ZF expression plasmid construction, Wolfe selection system testing, wiki coding and design, poster design

William Clerx
ZF target selection, wiki design, video editing, freelance wet lab, human practices

Sarah Choudhury
TolC selection system construction, TolC and Wolfe selection system testing, biobrick construction

Naomi Genuth
Majority of one-hybrid selection system construction and testing, biobrick construction and registration

Brandon Gerberich
ZF target selection, Wolfe selection system testing, interview cameraman and video editor, human practices

Mark Kopelman
Wet lab

Matt Lunati
TolC selection system construction and testing

Nida Naushad
Bioinformatics, chip sequences design and programming, ZF expression plasmid construction, chip library assembly, biobrick construction

Adviser Acknowledgments

Finally, we would like to specially acknowledge all of our teaching fellows (TFs) who have always been there for us, no matter how small or big the problem. Their advice has been of immeasurable value to our project's success. We would also like to also acknowledge the following TFs for their special contributions:

Daniel Goodman
Harvard iGEM logo design, wiki design (contributer to CSS and design of the navigation bar), and being an all around awesome team leader

Noah Taylor
Used lambda-red recombination to successfully change our zinc finger binding sites when MAGE was impractical, contributed a reverse translating function from his own research to bioinformatics

Srivatsan Raman
Collaborator in our project abstract and human practices page

References

1. Sriram Kosuri, Nikolai Eroshenko, Emily M LeProust, Michael Super, Jeffrey Way, Jin Billy Li, George M Church. (2010). Scalable gene synthesis by selective amplification of DNA pools from high-fidelity microchips. Nature Biotechnology, 28(12):1295-9. [http://www.nature.com/nbt/journal/v28/n12/full/nbt.1716.html]

2. Harris H. Wang, Farren J. Isaacs, Peter A. Carr, Zachary Z. Sun, George Xu, Craig R. Forest, George M. Church. Programming cells by multiplex genome engineering and accelerated evolution. (2009). Nature, 460(7257):894-8. [http://www.nature.com/nature/journal/v460/n7257/full/nature08187.html]

3. Yu D., H. M. Ellis, et al. (2000). An efficient recombination system for chromosome engineering in Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America 97(11): 5978-5983.[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC165854/]