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Fluorogenic label to quantify the cytosolic delivery of macromolecules.

ABSTRACT: The delivery of a macromolecule to the cytosol of human cells is assessed by using a pendant di-O-glycosylated derivative of fluorescein. Its fluorescence is unmasked by Escherichia coli?-galactosidase installed in the cytosol. Background is diminished by using RNAi to suppress the expression of GLB1, which encodes a lysosomal ?-galactosidase. This strategy was used to quantify the cytosolic entry of a highly cationic protein, ribonuclease A.

SUBMITTER: Chao TY

PROVIDER: S-EPMC3588161 | biostudies-literature | 2013 Mar

REPOSITORIES: biostudies-literature

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Fluorogenic label to quantify the cytosolic delivery of macromolecules.

Chao Tzu-Yuan TY Raines Ronald T RT

Molecular bioSystems 20130122 3

The delivery of a macromolecule to the cytosol of human cells is assessed by using a pendant di-O-glycosylated derivative of fluorescein. Its fluorescence is unmasked by Escherichia coliβ-galactosidase installed in the cytosol. Background is diminished by using RNAi to suppress the expression of GLB1, which encodes a lysosomal β-galactosidase. This strategy was used to quantify the cytosolic entry of a highly cationic protein, ribonuclease A. ...[more]

PMID: 23340874

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