Reza Faisal, Glazer Peter M
Departments of Therapeutic Radiology, Yale University School of Medicine, New Haven, CT, USA.
Methods Mol Biol. 2014;1114:115-42. doi: 10.1007/978-1-62703-761-7_8.
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.
通过外源性引入分子与内源性重组机制的相互作用，可以在体外和体内实现基因组靶向和编辑。前者包括一系列用于靶向诱导和利用该机制的序列特异性结合分子，例如分别通过三链体形成寡核苷酸（TFO）或三链体形成肽核酸（PNA）以及重组供体DNA。这种通过重组进行的通用靶向和编辑方法有助于实现高保真和低脱靶的基因组诱变、修复、表达和调控。在此，我们描述了三链体介导的基因组靶向和编辑的当前技术水平，并展望其潜在的转化和治疗应用。我们详细介绍了几种用于设计、递送和使用三链体形成分子和重组分子以介导和引入特定、可遗传且安全的基因组修饰的材料和方法。此外，我们指出了一些用于内源性基因组靶向和编辑位点鉴定的指导原则以及测试靶向和编辑效率的技术。
