Engineered Cys2His2 Zinc Finger Library for Efficient non -G Rich Target Selections

Engineered Cys₂His₂ zinc fingers for   efficient non-G rich Target selections

Princeton Docket # 12-2803

Researchers in the Lewis-Sigler Institute for Integrative Genomics, Princeton University have developed a novel zinc finger profile library, which provides improvements to and expands upon the existence of known, artificial zinc fingers.

Applications            

·         Use as artificial transcription factors for a given target within a genome of interest

·         Zinc Finger Nuclease - Site specific genome editing and repair

·         Targeting of chromatin modifying proteins throughput screening of compound libraries

·         Research tool to study gene function

Advantages  

·         Covers the vast majority of target sequences at 3 different registers of a protein

·         Expand the set of zinc fingers with known specificity to include CNN, ANN and TNN fingers.

·         Increases the number of engineered zinc fingers using alternative backbones

·         Gratuitous inducer

Zinc finger nucleases and recombinases have been used to make genomic modification in human cell lines and many model organisms. Zinc fingers are also used as artificial transcription factors that have been used to regulate specific genes within a genome of choice. This novel comprehensive set of zinc fingers could be used to assemble zinc finger arrays able to target auxiliary domains to almost any sequence of interest, thereby expanding the total number of targets that one may go after with this type of engineered protein. 

This library covers the vast majority of target sequences at 3 different registers of a protein.  This drastically increases the number of engineered zinc fingers outside of the common Zif268 finger 2 backbone, by engineering complete sets using finger 1 and finger 3. At all 3 positions/backbones we offer great improvement over previous data where there is significant enrichment of most fingers outside of the traditional G-rich context. Moreover, the described fingers at the N and C-terminal positions allows for the assembly of zinc fingers where the terminal fingers have been selected in the context under which they had been evolved.  Methods for zinc finger library assembly along with methods of assembling zinc finger arrays are also provided.

Zinc finger nucleases and recombinases have been used to make genomic modification in human cell lines and many model organisms. Zinc fingers are also used as artificial transcription factors that have been used to regulate specific genes within a genome of choice. This novel comprehensive set of zinc fingers could be used to assemble zinc finger arrays able to target auxiliary domains to almost any sequence of interest, thereby expanding the total number of targets that one may go after with this type of engineered protein. 

The Inventors

Marcus Noyes, PhD is an Associate Research Scholar, at the Lewis-Sigler Institute for Integrative Genomics, Department of Molecular Biology at Princeton University.

Mona Singh, PhD is a Professor of Computer Science and the Lewis-Sigler Institute for Integrative Genomic focusing on computational Biology.

Anton Persikov, PhD is a Postdoc in the Singh lab who focuses computational approaches to understanding zinc finger specificity.

Intellectual Property status and Commercialization Strategy

Patent protection is pending.

Princeton University¿s Office of Technology Licensing is interested in identifying appropriate partners for the further development and commercialization of this technology.

Contact:

Laurie Tzodikov

Princeton University Office of Technology Licensing ¿ (609) 258-7256¿ tzodikov@princeton.edu

Princeton docket # 12-2803