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Screening of cell‐penetrating peptides using mRNA display
Author(s) -
Lee JaeHun,
Song Hyun Seok,
Lee SunGu,
Park Tae Hyun,
Kim ByungGee
Publication year - 2012
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201100220
Subject(s) - cell penetrating peptide , hek 293 cells , hela , messenger rna , cell culture , cell , peptide , in vivo , microbiology and biotechnology , in vitro , embryonic stem cell , peptide library , chemistry , biology , peptide sequence , biochemistry , gene , genetics
Cell‐penetrating peptides (CPPs) are attractive vectors for in vivo and in vitro cellular uptake. Their use is, however, limited by insufficient understanding of their preference for a target cell. Here, a new CPP screening method is presented that uses mRNA display. After incubating the target cell lines, such as human embryonic kidney 293 (HEK 293) and HeLa cells, with an mRNA display library for 3 h at 37°C, the CPP‐mRNA nucleotide conjugates were harvested. These were amplified with PCR and subsequently sequenced. The screened CPPs for each cell line were identified after four rounds of selection. Among them, two peptides, MAMPGEPRRANVMAHKLEPASLQLR NSCA (CPPK) and MAPQRDTVGGRTTPPSWGPAKAQLRNSCA (CPPL) were selected, and the FITC‐labeled peptides were evaluated for their ability to penetrate cells. The screened CPPs were superior to polyarginine (R 11 ), which is widely used as a standard peptide and shows good cell penetration efficiency. Our method can be applied to other target cells for which CPPs have not yet been elucidated.