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A Method for the Acute and Rapid Degradation of Endogenous Proteins.

Cell | Dec 14, 2017

Methods for the targeted disruption of protein function have revolutionized science and greatly expedited the systematic characterization of genes. Two main approaches are currently used to disrupt protein function: DNA knockout and RNA interference, which act at the genome and mRNA level, respectively. A method that directly alters endogenous protein levels is currently not available. Here, we present Trim-Away, a technique to degrade endogenous proteins acutely in mammalian cells without prior modification of the genome or mRNA. Trim-Away harnesses the cellular protein degradation machinery to remove unmodified native proteins within minutes of application. This rapidity minimizes the risk that phenotypes are compensated and that secondary, non-specific defects accumulate over time. Because Trim-Away utilizes antibodies, it can be applied to a wide range of target proteins using off-the-shelf reagents. Trim-Away allows the study of protein function in diverse cell types, including non-dividing primary cells where genome- and RNA-targeting methods are limited.

Pubmed ID: 29153837 RIS Download

Mesh terms: Animals | Antibodies | Biochemistry | Protein Transport | Proteolysis

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Associated grants

  • Agency: Wellcome Trust, Id: MC_U105181010
  • Agency: Medical Research Council, Id: MC_U105192711
  • Agency: Medical Research Council, Id:

Addgene (Reagent, Plasmid)

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GE Healthcare

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ATCC

Global nonprofit biological resource center (BRC) and research organization that provides biological products, technical services and educational programs to private industry, government and academic organizations. Its mission is to acquire, authenticate, preserve, develop and distribute biological materials, information, technology, intellectual property and standards for the advancement and application of scientific knowledge. The primary purpose of ATCC is to use its resources and experience as a BRC to become the world leader in standard biological reference materials management, intellectual property resource management and translational research as applied to biomaterial development, standardization and certification. ATCC characterizes cell lines, bacteria, viruses, fungi and protozoa, as well as develops and evaluates assays and techniques for validating research resources and preserving and distributing biological materials to the public and private sector research communities.

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Addgene

A non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. They work with laboratories to assemble a high-quality library of published and useful plasmids and their associated cloning/sequence data for use in research and discovery. By linking plasmids with articles, scientists can always find data related to the materials they request. There is no cost to deposit plasmids to Addgene and it will store samples in triplicate (including one at an offsite backup facility), sequence key regions for validation, and handle the appropriate Material Transfer Agreements (MTAs) with institutions. Additionally, users can create a webpage that directs scientists to request plasmids. Material Transfer Agreements (MTAs) allow open exchange to occur because they offer intellectual property and liability protection for material providers. Institutions that have deposited materials at Addgene require a MTA for each transfer of material.

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Novus Biologicals

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Jackson Laboratory

An independent, nonprofit organization focused on mammalian genetics research to advance human health. Their mission is to discover the genetic basis for preventing, treating, and curing human disease, and to enable research for the global biomedical community. Jackson Laboratory breeds and manages colonies of mice as resources for other research institutions and laboratories, along with providing software and techniques. Jackson Lab also conducts genetic research and provides educational material for various educational levels.

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Imaris

Interactive Vision In The Life SciencesCombining speed, precision and ease-of-use, Imaris provides an impressive range of capabilities for working with three-dimensional images. Move easily between simple and advanced 3D views. Use flexible editing and processing functions, such as interactive surface rendering and object slicing capabilities. And output to standard TIFF, Quicktime and AVI formats. Imaris accepts virtually all image formats that are used in confocal microscopy and many of those used in wide-field image acquisition.BitPlane, Inc. also releases the following companion products:- ImarisXT is a multi-functional two-way interface from Imaris to both classic programming languages and to Matlab.- Imaris MeasurementPro and Imaris Track provide the ability to segment and follow the movement of independent objects in a 3D image over time. Objects can either be Surface Objects or Spots, created automatically or manually.- ImarisColoc enables users to easily isolate, visualize, and quantify regional overlap in 3D and 4D images and automates the process of selecting colocalized image areas.

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