RT Journal Article
SR Electronic
T1 Eukaryotically expressed encapsulins as orthogonal compartments for multiscale molecular imaging
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 222083
DO 10.1101/222083
A1 Felix Sigmund
A1 Christoph Massner
A1 Philipp Erdmann
A1 Anja Stelzl
A1 Hannes Rolbieski
A1 Helmut Fuchs
A1 Martin Hrabé de Angelis
A1 Mitul Desai
A1 Sarah Bricault
A1 Alan Jasanoff
A1 Vasilis Ntziachristos
A1 Jüergen Plitzko
A1 Gil G. Westmeyer
YR 2017
UL http://biorxiv.org/content/early/2017/11/27/222083.abstract
AB We have genetically controlled compartmentalization in eukaryotic cells by heterologous expression of bacterial encapsulin shell and cargo proteins to engineer enclosed enzymatic reactions and size-controlled metal biomineralization. The orthogonal shell protein (EncA) from M. xanthus efficiently auto-assembled inside mammalian cells into nanocompartments to which sets of native (EncB,C,D) and engineered cargo proteins self-targeted. This enabled localized bimolecular fluorescence and enzyme complementation with selective access to substrates via the pores in the nanoshell. Encapsulation of the enzyme tyrosinase lead to the confinement of toxic melanin production for robust detection via multispectral optoacoustic tomography (MSOT). Co-expression of ferritin-like native cargo (EncB or EncC) resulted in efficient iron sequestration that produced substantial contrast by magnetic resonance imaging (MRI) and enabled magnetic cell sorting. The monodisperse, spherical, and iron-loading nanoshells also proved to be excellent genetically encoded markers for cryo-electron tomography (cryo-ET). 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 therapy.