Construction and Screening of a Lentiviral Secretome Library

Tao Liu, Panpan Jia, Huailei Ma, Sean A. Reed, Xiaozhou Luo, H. Benjamin Larman, Peter G. Schultz

Research output: Contribution to journalArticlepeer-review

Abstract

Over 2,000 human proteins are predicted to be secreted, but the biological function of the many of these proteins is still unknown. Moreover, a number of these proteins may act as new therapeutic agents or be targets for the development of therapeutic antibodies. To further explore the extracellular proteome, we have developed a secretome-enriched open reading frame (ORF) library that can be readily screened for autocrine activity in cell-based phenotypic or reporter assays. Next-generation sequencing (NGS) and database analysis predict that the library contains approximately 900 ORFs encoding known secreted proteins (accounting for 77.8% of the library), as well as genes encoding potentially unknown secreted proteins. In a proof-of-principle study, human TF-1 cells were screened for proliferative factors, and the known cytokine GMCSF was identified as a dominant hit. This library offers a relatively low-cost and straightforward approach for functional autocrine screens of secreted proteins.

Original languageEnglish (US)
Pages (from-to)767-771.e3
JournalCell Chemical Biology
Volume24
Issue number6
DOIs
StatePublished - Jun 22 2017

Keywords

  • autocrine
  • lentivirus
  • next-generation sequencing
  • open reading frame
  • secretome

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

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