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Analysis of the cellular uptake of a novel cell penetrating peptide and peptoid
Author(s) -
Pietak MariaClaire,
Chaput Dale I.,
Zamore Elizabeth,
Goldberg Bradley,
Danowski Barbara,
Kehlbeck Joanne
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.895.2
Subject(s) - cell penetrating peptide , transfection , intracellular , endocytosis , chemistry , cell , peptide , cell culture , enzyme , substrate (aquarium) , biophysics , microbiology and biotechnology , biochemistry , biology , ecology , genetics , gene
Intracellular drug delivery is restricted by the selective permeability of the plasma membrane. This can be overcome with cell penetrating peptides (CPPs) which are short peptide sequences that can traverse the cell membrane with attached cargo. Here, we investigate the uptake of a novel CPP, thought to be more permeable due to increased polarizability, by employing an enzyme substrate assay with a fluorescence read‐out. Swiss 3t3 and Madin Darby Canine Kidney (MDCK) cells will be transfected with an enzyme. The experimental CPP, along with positive and negative controls, will be conjugated to the substrate for the enzyme, and incubated with live, transfected cells. The ideal substrate (the 'cargo') will only fluoresce once inside the cell, allowing us to compare uptake efficiencies. We will be using endocytosis inhibitors to identify the mechanism of uptake. Thus far, we have optimized DNA transfection in both cell lines by comparison of transfection reagents, cell confluence, and incubation time. Next, we will focus on perfecting the enzyme detection assay and evaluating our CPPs. Supported by Union College IEF, Merck/AAAS Summer Fellowship, and Sciortino Summer Fellowship, Union College.