Selective gene silencing in activated leukocytes by targeting siRNAs to the integrin lymphocyte function-associated antigen-1

Silencing gene expression by RNAi is a powerful method for exploring gene function and validating drug targets and potentially for therapy. Lymphocytes and other primary blood cells are resistant to lipid-based transfection in vitro and are difficult to target in vivo. We show here that antibody-pro...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2007-03, Vol.104 (10), p.4095-4100
Main Authors: Peer, Dan, Zhu, Pengcheng, Carman, Christopher V, Lieberman, Judy, Shimaoka, Motomu
Format: Article
Language:English
Subjects:
Animals
Antibodies
Antigens, Nuclear - biosynthesis
Biological Sciences
Cell Adhesion
Cell adhesion & migration
Cell Proliferation
Cyclins
DNA-Binding Proteins - biosynthesis
Dose-Response Relationship, Drug
Drug therapy
Gene expression
Gene Silencing
Humans
Immune system
K562 Cells
Ku Autoantigen
Leukocytes
Lymphocyte Function-Associated Antigen-1 - metabolism
Lymphocyte Function-Associated Antigen-1 - physiology
Lymphocytes
Mice
Mice, SCID
Protamines
Proteins
Receptors, CCR5 - metabolism
Recombinant Fusion Proteins - chemistry
RNA, Small Interfering - metabolism
Rodents
Small interfering RNA
T lymphocytes
T-Lymphocytes - metabolism
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Summary:Silencing gene expression by RNAi is a powerful method for exploring gene function and validating drug targets and potentially for therapy. Lymphocytes and other primary blood cells are resistant to lipid-based transfection in vitro and are difficult to target in vivo. We show here that antibody-protamine fusion proteins targeting the human integrin lymphocyte function-associated antigen-1 (LFA-1) efficiently deliver siRNAs and specifically induce silencing in primary lymphocytes, monocytes, and dendritic cells. Moreover, a fusion protein constructed from an antibody that preferentially recognizes activation-dependent conformational changes in LFA-1 selectively targets activated leukocytes and can be used to suppress gene expression and cell proliferation only in activated lymphocytes. The siRNA-fusion protein complexes do not cause lymphocyte activation or induce IFN responses. K562 cells expressing latent WT or constitutively activated LFA-1 engrafted in the lungs of SCID mice are selectively targeted by intravenously injected fusion protein-siRNA complexes, demonstrating the potential in vivo applicability of LFA-1-directed siRNA delivery.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0608491104