Tunable light and drug induced depletion of target proteins

Autor: Jack A. Bates, Michael D. Bartoschek, Wen Deng, Hai Wei, Barbara Conradt, Heinrich Leonhardt
Jazyk: angličtina
Rok vydání: 2020
Předmět:
0301 basic medicine
DNA Repair
Light
General Physics and Astronomy
Apoptosis
law.invention
DNA Ligase ATP
0302 clinical medicine
Ubiquitin
law
lcsh:Science
Cell Engineering
Caenorhabditis elegans
Regulation of gene expression
Multidisciplinary
biology
Chemistry
Biological techniques
Gene Expression Regulation
Developmental

Lamin Type A
Cell biology
DNA-Binding Proteins
DARPin
Caspases
Recombinant DNA
DNA repair
DNA damage
Ubiquitin-Protein Ligases
Science
Cytological Techniques
Green Fluorescent Proteins
Protein degradation
General Biochemistry
Genetics and Molecular Biology

Article
03 medical and health sciences
Proliferating Cell Nuclear Antigen
Developmental biology
Animals
Humans
Caenorhabditis elegans Proteins
fungi
General Chemistry
biology.organism_classification
030104 developmental biology
biology.protein
lcsh:Q
030217 neurology & neurosurgery
DNA Damage
HeLa Cells
Transcription Factors
Zdroj: Nature Communications, Vol 11, Iss 1, Pp 1-13 (2020)
Nature Communications
ISSN: 2041-1723
Popis: Biological processes in development and disease are controlled by the abundance, localization and modification of cellular proteins. We have developed versatile tools based on recombinant E3 ubiquitin ligases that are controlled by light or drug induced heterodimerization for nanobody or DARPin targeted depletion of endogenous proteins in cells and organisms. We use this rapid, tunable and reversible protein depletion for functional studies of essential proteins like PCNA in DNA repair and to investigate the role of CED-3 in apoptosis during Caenorhabditis elegans development. These independent tools can be combined for spatial and temporal depletion of different sets of proteins, can help to distinguish immediate cellular responses from long-term adaptation effects and can facilitate the exploration of complex networks.
Methods to control protein degradation usually require prior genetic engineering of each specific protein of interest. Here the authors present a toolbox for targeted depletion of endogenous proteins in cells and organisms, which is based on light- and drug-controlled recombinant E3 ubiquitin ligases.
Databáze: OpenAIRE