Receptor-mediated delivery of engineered nucleases for genome modification.
Nucleic Acids Res
; 41(19): e182, 2013 Oct.
Article
em En
| MEDLINE
| ID: mdl-23956220
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Receptores da Transferrina
/
Desoxirribonucleases
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Reparo Gênico Alvo-Dirigido
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article