Abstract:
: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.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Chen Z,Jaafar L,Agyekum DG,Xiao H,Wade MF,Kumaran RI,Spector DL,Bao G,Porteus MH,Dynan WS,Meiler SEdoi
10.1093/nar/gkt710subject
Has Abstractpub_date
2013-10-01 00:00:00pages
e182issue
19eissn
0305-1048issn
1362-4962pii
gkt710journal_volume
41pub_type
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