Laboratory of endoplasmic reticulum (ER) quality control and protein misfolding diseases

Lab Description
Main areas of interest are in the mechanisms of protein folding and trafficking in the early secretory pathway of mammalian cells and the development of ER stress upon their malfunction, particularly in neurodegenerative diseases.

Specific research topics are:
• Protein misfolding diseases, including neurodegenerative diseases.
• ER stress.
• Mechanisms of endoplasmic reticulum (ER) protein folding and quality control.
• Sugar chain trimming processes as cellular signals for glycoprotein folding status.
• Delivery of misfolded proteins to ER-associated degradation (ERAD).
• Subcellular compartmentalization of ER quality control and ERAD.

Technology
• Biochemical and imaging studies in mammalian cells.
• Cytotoxicity and apoptosis assays.
• Subcellular fractionation, density gradients.
• Protein analysis: Western blot, immunoprecipitation, folding and glycosylation analysis. Protein interactions, coimmunoprecipitation.
• Radioactive pulse chase analysis.
• DNA cloning and engineering of constructs, mutants, fluorescent fusion proteins.
• RNA analysis, qPCR.
• Gene expression by cell transfection and infection, retrovirus and lentivirus vectors. Stable cell line selection.
• SiRNA and shRNA knockdown. Caspr/Cas9.
• Subcellular localization and dynamics. Immunofluorescence, confocal and live cell imaging. FRET, FRAP, FLIP.
Available Research Services
1. Analysis of all ER stress pathways and their modulation.
2. Cytotoxicity and apoptosis assays.
3. Protein subcellular localization and dynamics in fixed and live cells.
4. Analysis of protein folding, glycosylation, turnover rate, trafficking.
Lab facilities
The laboratory is 60 m2 and it has all the small equipment required for molecular biology and biochemical techniques, including equipment for electrophoresis, gel and immunoblot imager, PCR thermocycler, ELISA reader, incubators, water baths, microfuges, etc. For tissue culture we have a separate suite with two sterile hoods, CO2 incubators and a liquid nitrogen locator for our own use. The department and the Faculty of Life Sciences are well equipped with large equipment, refrigerated centrifuges, ultracentrifuges, -80˚C freezers, shaker-incubators, lyophilizers, fluorescence, confocal and electron microscopes, phosphorimager, etc. This equipment is shared. The imaging unit of Tel Aviv University has further shared specialized microscopes, including confocal microscopes and STED. The personnel in the laboratory include a postdoctoral fellow, two PhD students, two MSc students and a research assistant.

Potential industries
1. Pharma
2. Biotech

Contact Information
Prof. Gerardo Lederkremer
Office: 972-3-640-9239
Lab: 972-3-640-7610
e-mail:gerardo@post.tau.ac.il
website: The Lederkremer Lab Home Page

Project manager

avi Wener
Research services

Project researchers

Gerardo Z. (Zelmar) Lederkremer
T.A.U Tel Aviv University, Life Sciences
Cell Research and Immunology