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Triplex-Mediated Genome Targeting and Editing

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Gene Correction

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1114))

Abstract

Genome targeting and editing in vitro and in vivo can be achieved through an interplay of exogenously introduced molecules and the induction of endogenous recombination machinery. The former includes a repertoire of sequence-specific binding molecules for targeted induction and appropriation of this machinery, such as by triplex-forming oligonucleotides (TFOs) or triplex-forming peptide nucleic acids (PNAs) and recombinagenic donor DNA, respectively. This versatile targeting and editing via recombination approach facilitates high-fidelity and low-off-target genome mutagenesis, repair, expression, and regulation.

Herein, we describe the current state-of-the-art in triplex-mediated genome targeting and editing with a perspective towards potential translational and therapeutic applications. We detail several materials and methods for the design, delivery, and use of triplex-forming and recombinagenic molecules for mediating and introducing specific, heritable, and safe genomic modifications. Furthermore we denote some guidelines for endogenous genome targeting and editing site identification and techniques to test targeting and editing efficiency.

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Acknowledgments

We gratefully acknowledge members of the Glazer Laboratory for helpful discussions. This work was supported by a National Institutes of Health (NIH) grant R01HL082655 and by a Doris Duke Innovations in Clinical Research Award (to P.M.G.). A National Institute of Diabetes and Digestive and Kidney Diseases Experimental and Human Pathobiology Postdoctoral Fellowship from NIH grant T32DK007556 also provided support (to F.R.). The authors declared no conflict of interest.

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Authors and Affiliations

  1. Departments of Therapeutic Radiology, Yale University School of Medicine, New Haven, CT, USA

    Faisal Reza

  2. Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, CT, USA

    Peter M. Glazer

  3. Department of Genetics, Yale University School of Medicine, New Haven, CT, USA

    Peter M. Glazer

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  1. Faisal Reza
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  2. Peter M. Glazer
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Editors and Affiliations

  1. Georgia Institute of Technology, Atlanta, Georgia, USA

    Francesca Storici

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Reza, F., Glazer, P.M. (2014). Triplex-Mediated Genome Targeting and Editing. In: Storici, F. (eds) Gene Correction. Methods in Molecular Biology, vol 1114. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-761-7_8

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  • DOI: https://doi.org/10.1007/978-1-62703-761-7_8

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-760-0

  • Online ISBN: 978-1-62703-761-7

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