Massachusetts Institute of Technology

Carl O Pabo

Jin-Soo Kim

Phillip A Sharp

Joel L Pomerantz

J Keith Joung

Jeffrey N Miller

Edward J Rebar

Bryan S Wang

Harvey A Greisman

Jin-Soo-Samsung Biomedical Research Institute Kim

The invention provides nonnaturally occurring dimerizing peptides, and methods for their production. Such peptides are useful to mediate association of linked functional proteins domains. In particular, such peptides are useful for mediating association of complexes of multiple zinc finger proteins thereby affording greater specificity and/or affinity in binding of the zinc finger proteins to proximately spaced target segments.

Dimerizing peptides

Polynucleotides encoding chimeric proteins, and methods for their production and use are disclosed. The chimeric proteins comprise a flexible linker between two zinc finger DNA-binding domains, wherein the linker contains eight or more amino acids between the second conserved histidine residue of the carboxy-terminal zinc finger of the first domain and the first conserved cysteine residue of the amino-terminal zinc finger of the second domain.

Poly zinc finger proteins with improved linkers

Chimeric proteins containing composite DNA-binding regions are disclosed together with DNA constructs encoding them, compositions containing them and applications in which they are useful.

Chimeric DNA-binding proteins

The present invention provides methods and compositions for interaction trap assays for detecting protein-protein, protein-DNA, or protein-RNA interactions. The methods and compositions of the invention may also be used to identify agents which may agonize or antagonize a protein-protein, protein-DNA, or protein-RNA interaction. In certain embodiments, the interaction trap system of the invention is useful for screening libraries with greater than 10members. In other embodiments, the interaction trap system of the invention is used in conjunction with flow cytometry. The invention further provides a means for simultaneously screening a target protein or nucleic acid sequence for the ability to interact with two or more test proteins or nucleic acids.

Methods and compositions for interaction trap assays

The present invention provides methods and compositions for interaction trap assays for detecting protein-protein, protein-DNA, or protein-RNA interactions. The methods and compositions of the invention may also be used to identify agents which may agonize or antagonize a protein-protein, protein-DNA, or protein-RNA interactions. In certain embodiments, the interaction trap system of the invention is useful for screening libraries with greater than 10members. In other embodiments, the interaction trap system of the invention is used in conjunction with flow cytometry. The invention further provides a means for simultaneously screening a target protein or nucleic acid sequence for the ability to interact with two or more test proteins or nucleic acids.

Nucleic acid encoding poly-zinc finger proteins with improved linkers

Chimeric proteins, and methods for their production and use are disclosed. The chimeric proteins comprise a flexible linker between two zinc finger DNA-binding domains, wherein the linker comprises eight or more amino acids between the second conserved histidine residue of the carboxy-terminal zinc finger of the first domain and the first conserved cysteine residue of the amino-terminal zinc finger of the second domain.

Provided herein are chimeric zinc finger proteins comprising flexible linkers of at least 6 amino acids in length. Also provided are methods of making and using these chimeric zinc finger proteins.

The present invention provides methods for making zinc finger proteins with high affinity for their targets, by sequentially selecting each finger of the protein.

General strategy for selecting high-affinity zinc finger proteins for diverse DNA target sites

Chineric proteins containing composite DNA-binding regions are disclosed together with DNA constructs encoding them, compositions containing them and applications in which they are useful.

Described is a polypeptide comprising one or more zinc fingers. The polypeptide binds to new polynucleotide subsites with high affinity and consequently has a binding specificity that differs from wild type zinc finger proteins. The binding occurs through contacts between certain amino acid residues of the zinc fingers and the nucleic acids of the subsites. The polypeptide sequence of at least one zinc finger differs from wild type zinc fingers, and the difference involves at least one amino acid residue that contacts the bases of the polynucleotide during binding.

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Newton, MA, United States of America

Carl O Pabo