title: Screening of cell-penetrating peptides using mRNA display.
abstract: 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.
authors.affiliation: Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University, Seoul, South Korea.
authors.forename: Jae-Hun
authors.initials: JH
authors.lastname: Lee
authors.forename: Hyun Seok
authors.initials: HS
authors.lastname: Song
authors.forename: Tai Hyun
authors.initials: TH
authors.lastname: Park
authors.forename: Tae Hyun
authors.initials: TH
authors.lastname: Park
authors.forename: Sun-Gu
authors.initials: SG
authors.lastname: Lee
authors.forename: Byung-Gee
authors.initials: BG
authors.lastname: Kim
mesh_terms: D057846:Cell-Penetrating Peptides / Q000235:genetics* / Q000302:isolation & purification*; D016650:Fluorescein-5-isothiocyanate; D005822:Genetic Vectors; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics
chemical_list: D057846:Cell-Penetrating Peptides; D012333:RNA, Messenger; D016650:Fluorescein-5-isothiocyanate