In Vivo Genome Editing as a Therapeutic Approach

Abstract

Genome editing has been well established as a genome engineering tool that enables researchers to establish causal linkages between genetic mutation and biological phenotypes, providing further understanding of the genetic manifestation of many debilitating diseases. More recently, the paradigm of genome editing technologies has evolved to include the correction of mutations that cause diseases via the use of nucleases such as zinc-finger nucleases (ZFN), transcription activator-like effector nucleases (TALENs), and more recently, Cas9 nuclease. With the aim of reversing disease phenotypes, which arise from somatic gene mutations, current research focuses on the clinical translatability of correcting human genetic diseases in vivo, to provide long-term therapeutic benefits and potentially circumvent the limitations of in vivo cell replacement therapy. In this review, in addition to providing an overview of the various genome editing techniques available, we have also summarized several in vivo genome engineering strategies that have successfully demonstrated disease correction via in vivo genome editing. The various benefits and challenges faced in applying in vivo genome editing in humans will also be discussed.

Novel HSP90-PI3K Dual Inhibitor Suppresses Melanoma Cell Proliferation by Interfering with HSP90-EGFR Interaction and Downstream Signaling Pathways

“…The malignant melanoma cell lines, including SK-MEL-28, A875, B16, A375, and A2058 were obtained from the Chinese Center for Type Culture Collection (Wuhan, China) and cultured in DMEM (Dulbecco's modified Eagle's medium) with 10% FBS (fetal bovine serum) and streptomycin. The cytotoxicity assay was performed with the MTT method, as previously described [69][70][71].…”

Gene therapy trials for haemophilia: a step closer to a cure?

“…lentivirus) and non-viral approaches to gene delivery are under development (reviewed in [30,31]). Studies using ex-vivo gene editing, including zinc-finger nuclease (ZFN) and CRISPR/ Cas9 systems are also ongoing [32]. We are undoubtedly now one step closer but not at the end of the road.…”

Mitigating off-target effects in CRISPR/Cas9-mediated in vivo gene editing

“…These include, but are not limited to, zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 endonucleases. Of the three, ZFN and TALEN are nucleases containing a customisable DNA-binding domain, which is commonly fused to a FokI DNA-cleavage domain [2][3][4]. The CRISPR/Cas9 system consists of a DNA-cleaving endonuclease (Cas9) associated with a guide RNA that recognises and binds to the targeted sequences [5][6][7].…”

“…Similarly, significant rates of OTEs had been reported in in vitro CRISPR/Cas9 edited human cell lines [18][19][20][21][22] and non-viable human embryos [23]. The CRISPR/Cas9 gene editing technique holds many advantages over ZFNs and TALENs in targeting disorders with a distinct genetic aetiology-the ease of designing the guide RNA sequences, the computational determination of OTEs based on genomic sequences with high similarity to the target locus (first-order sequence screens) [24][25][26][27] and the possibility of editing multiple genomic loci simultaneously [4]. Nonetheless, it remains prudent for clinical applications of CRISPR/Cas9 to implement constructs that can minimise OTEs.…”