Screening of cell-penetrating peptides using mRNA display

Jae Hun Lee, Hyun Seok Song, Sun Gu Lee, Tae Hyun Park, Byung Gee Kim

Research output: Contribution to journalArticlepeer-review

13 Scopus citations


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.

Original languageEnglish
Pages (from-to)387-396
Number of pages10
JournalBiotechnology Journal
Issue number3
StatePublished - Mar 2012


  • Cell-penetrating peptide
  • MRNA display
  • Peptide screening


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