Receptor-mediated delivery of engineered nucleases for genome modification

Zhong Chen, Lahcen Jaafar, Davies G. Agyekum, Haiyan Xiao, Marlene F. Wade, R. Ileng Kumaran, David L. Spector, Gang Bao, Matthew H. Porteus, William S. Dynan, Steffen E Meiler

Research output: Contribution to journalArticlepeer-review

35 Scopus citations


Engineered nucleases, which incise the genome at predetermined sites, have a number of laboratory and clinical applications. There is, however, a need for better methods for controlled intracellular delivery of nucleases. Here, we demonstrate a method for ligand-mediated delivery of zinc finger nucleases (ZFN) proteins using transferrin receptor-mediated endocytosis. Uptake is rapid and efficient in established mammalian cell lines and in primary cells, including mouse and human hematopoietic stem-progenitor cell populations. In contrast to cDNA expression, ZFN protein levels decline rapidly following internalization, affording better temporal control of nuclease activity. We show that transferrin-mediated ZFN uptake leads to site-specific in situ cleavage of the target locus. Additionally, despite the much shorter duration of ZFN activity, the efficiency of gene correction approaches that seen with cDNA-mediated expression. The approach is flexible and general, with the potential for extension to other targeting ligands and nuclease architectures.

Original languageEnglish (US)
Pages (from-to)e182
JournalNucleic Acids Research
Issue number19
StatePublished - Oct 2013

ASJC Scopus subject areas

  • Genetics


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