6XWY

Highly pH-resistant long stokes-shift, red fluorescent protein mCRISPRed

Summary for 6XWY

• Entry DOI: 10.2210/pdb6xwy/pdb
• Descriptor: Red fluorescent protein eqFP611, 1,2-ETHANEDIOL, MALONIC ACID, ... (4 entities in total)
• Functional Keywords: ph resistant, long stokes shift, lss, red fluorescent protein, fluorescent protein
• Biological source: Entacmaea quadricolor (Bubble-tip anemone)
• Total number of polymer chains: 4
• Total formula weight: 99766.82

Authors

Erdogan, M.,Fabritius, A.,Basquin, J.,Griesbeck, O. (deposition date: 2020-01-24, release date: 2020-02-05, Last modification date: 2024-01-24)

Primary citation

Erdogan, M.,Fabritius, A.,Basquin, J.,Griesbeck, O.
Targeted In Situ Protein Diversification and Intra-organelle Validation in Mammalian Cells.
Cell Chem Biol, 27:610-621.e5, 2020

PubMed Abstract: Engineered proteins must be phenotypically selected for function in the appropriate physiological context. Here, we present a versatile approach that allows generating panels of mammalian cells that express diversified heterologous protein libraries in the cytosol or subcellular compartments under stable conditions and in a single-variant-per-cell manner. To this end we adapt CRISPR/Cas9 editing technology to diversify targeted stretches of a protein of interest in situ. We demonstrate the utility of the approach by in situ engineering and intra-lysosome specific selection of an extremely pH-resistant long Stokes shift red fluorescent protein variant. Tailoring properties to specific conditions of cellular sub-compartments or organelles of mammalian cells can be an important asset to optimize various proteins, protein-based tools, and biosensors for distinct functions.

PubMed: 32142629
DOI: 10.1016/j.chembiol.2020.02.004

Experimental method

X-RAY DIFFRACTION (1.75 Å)