Intracellular targeting of endogenous proteins
AssigneeDana-Farber Cancer Institute, Inc.
InventorsWayne A. Marasco, Jennifer Richardson
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Abstract
The present invention relates to a method by which one can target an undesired target molecule or target antigen, preferably an endogenous protein. The method comprises the intracellular expression of an antibody capable of binding to the target. A DNA sequence is delivered to a cell, the DNA sequence contains a sufficient number of nucleotides coding for the portion of an antibody capable of binding to the target operably linked to a promoter that will permit expression of the antibody in the cell(s) of interest. The antibody is then expressed intracellularly and binds to the target, thereby disrupting the target from its normal actions.
Claims32
A vector for the intracellular binding of an endogenous protein that traverses a cellular membrane, referred to as a target antigen, which comprises:
- a nucleic acid segment encoding a variable heavy chain sequence of an antibody which selectively binds to said target antigen and said antibody's corresponding variable light chain sequence operably linked to a promoter, wherein said nucleic acid sequence does not encode said antibody's wild-type leader sequence for the variable chains and said antibody's secretory sequences,
- thereby adapting said antibody expressed by said nucleic acid sequence for intracellular binding to said target antigen.
A DNA vector comprising a gene encoding an antibody that binds to an IL2R.alpha. operably linked to a promoter, wherein the antibody gene does not encode a secretory signal sequence.
A vector for the intracellular binding of an endogenous protein that traverses a cellular membrane, referred to as a target antigen, which comprises:
- a nucleic acid segment encoding a variable heavy chain sequence of an antibody which selectively binds to said target antigen and said antibody's corresponding variable light chain sequence operably linked to a promoter, wherein said nucleic acid sequence also encodes an endoplasmic reticulum or golgi apparatus localization sequence,
- thereby adapting said antibody expressed by said nucleic acid sequence to be retained in the endoplasmic reticulum-golgi apparatus for intracellular binding to said target antigen.
A method for the intracellular binding of a protein that traverses a cellular membrane, referred to as a target antigen, which comprises:
- (a) delivery of a nucleic acid segment containing a promoter operably linked to a gene encoding an antibody that binds to said target antigen, wherein the antibody expressed by said gene does not have said antibody's secretory sequences or any substitution for said secretory sequences;
- (b) intracellular expression of said antibody in a form that binds to said target antigen;
- and
- (c) intracellular binding of said target antigen by said antibody.
A method for the intracellular binding of a protein that traverses a cellular membrane, referred to as a target antigen, which comprises:
- (a) delivery of a nucleic acid segment containing a promoter operably linked to an antibody gene encoding an antibody that binds to said target antigen, wherein the nucleic acid segment also encodes an endoplasmic reticulum or golgi apparatus localization sequence;
- (b) intracellular expression of said antibody in a form that binds to said target antigen;
- (c) localization of said antibody in said cell's endoplasmic reticulum golgi apparatus;
- and
- (d) intracellular binding of said target antigen by said antibody.
Dependencies are inferred from the claim text.