Bacterial encapsulins as orthogonal compartments for mammalian cell engineering

Nature Communications
Felix SigmundGil G Westmeyer

Abstract

We genetically controlled compartmentalization in eukaryotic cells by heterologous expression of bacterial encapsulin shell and cargo proteins to engineer enclosed enzymatic reactions and size-constrained metal biomineralization. The shell protein (EncA) from Myxococcus xanthus auto-assembles into nanocompartments inside mammalian cells to which sets of native (EncB,C,D) and engineered cargo proteins self-target enabling localized bimolecular fluorescence and enzyme complementation. Encapsulation of the enzyme tyrosinase leads to the confinement of toxic melanin production for robust detection via multispectral optoacoustic tomography (MSOT). Co-expression of ferritin-like native cargo (EncB,C) results in efficient iron sequestration producing substantial contrast by magnetic resonance imaging (MRI) and allowing for magnetic cell sorting. The monodisperse, spherical, and iron-loading nanoshells are also excellent genetically encoded reporters for electron microscopy (EM). In general, eukaryotically expressed encapsulins enable cellular engineering of spatially confined multicomponent processes with versatile applications in multiscale molecular imaging, as well as intriguing implications for metabolic engineering and cellular t...Continue Reading

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Sep 13, 2020·Biotechnology and Bioengineering·Jesse A Jones, Tobias W Giessen
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Datasets Mentioned

BETA
EMD-5917

Methods Mentioned

BETA
electron microscopy
co-immunoprecipitation
fluorescence imaging
confocal microscopy
transfection
Co-IP
transmission electron microscopy
electrophoresis
chips

Software Mentioned

DYNAMICS
Python
ImageJ
Matlab
ViewMSOT
Image Sequence Analysis Tool
MassLynx

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