Zinc-Finger Nucleases for Somatic Gene Therapy: The Next Frontier

Abstract

Zinc-finger nucleases (ZFNs) are a powerful tool that can be used to edit the human genome ad libitum. The technology has experienced remarkable development in the last few years with regard to both the target site specificity and the engineering platforms used to generate zinc-finger proteins. As a result, two phase I clinical trials aimed at knocking out the CCR5 receptor in T cells isolated from HIV patients to protect these lymphocytes from infection with the virus have been initiated. Moreover, ZFNs have been successfully employed to knockout or correct disease-related genes in human stem cells, including hematopoietic precursor cells and induced pluripotent stem cells. Targeted genome engineering approaches in multipotent and pluripotent stem cells hold great promise for future strategies geared toward correcting inborn mutations for personalized cell replacement therapies. This review describes how ZFNs have been applied to models of gene therapy, discusses the opportunities and the risks associated with this novel technology, and suggests future directions for their safe application in therapeutic genome engineering.

Computational Tools for Investigating RNA-Protein Interaction Partners

“…Finally, another important future direction in research on RNAprotein interactions is the rational design of RNA-protein interfaces. Engineered DNA binding proteins, such as ZFNs and TALENS, have become enormously powerful tools for genome engineering, and are poised to enter clinical settings [56][57][58]. Likewise, RNA-binding proteins engineered to recognize specific RNA sequences [36] could become valuable tools for manipulating post-transcriptional regulatory networks in the research laboratory, and potentially, important therapeutic agents for treating genetic and infectious diseases.…”

The Nontoxic Cell Cycle Modulator Indirubin Augments Transduction of Adeno-Associated Viral Vectors and Zinc-Finger Nuclease-Mediated Gene Targeting

“…Meanwhile, this technology has been increasingly employed in biotechnology, and therapeutic applications are tangible, as epitomized by the introduction of targeted alterations in the genome of multi-and pluripotent human stem cells (Chamberlain et al, 2004;Hockemeyer et al, 2009;Zou et al, 2009;Benabdallah et al, 2010). Two major platforms have been described to overcome the low HR frequency in such cells: a viral vector platform based on adeno-associated virus (AAV) (Chamberlain et al, 2004;Mitsui et al, 2009;Khan et al, 2010) and DNA double-strand break (DSB)-inducing agents, such as meganucleases (Silva et al, 2011;Takeuchi et al, 2011), zinc-finger nucleases (ZFNs) (Urnov et al, 2010;Carroll, 2011;Rahman et al, 2011), and transcriptional activator-like effector (TALE) nucleases (Bogdanove and Voytas, 2011;Mussolino and Cathomen, 2012).…”

The Pharmacology of Regenerative Medicine

“…In the case of genetic disorders, it should be possible to correct mutations prior to returning cells to the patient (Xu et al, 2009;Kazuki et al, 2010;Khan et al, 2010;Howden et al, 2011;Wong and Chiu, 2011). For patients with chronic infections such as a hepatitis or immunodeficiency virus, it may be possible to arm cells with protective antiviral genetic elements (Kamata et al, 2010;Rahman et al, 2011).…”