Induced Pluripotency and Gene Editing in Disease Modelling: Perspectives and Challenges

Seah, Seng Hong; Farran, Chadi A. El; Warrier, Tushar; Xu, Jian; Loh, Yuin-Han

doi:10.3390/ijms161226119

Cited by 19 publications

(17 citation statements)

References 148 publications

(188 reference statements)

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“…Previous studies have reported the application of the genome-editing technique in the human pluripotent stem cells to model diseases that were caused by single-gene mutations for drug discovery [30,31]. In the present study, we generated GLA-null HEK-293T cell lines by CRISPR/Cas9 genome-editing system.…”

Section: Discussionmentioning

confidence: 97%

Using CRISPR/Cas9-Mediated GLA Gene Knockout as an In Vitro Drug Screening Model for Fabry Disease

Song¹,

Chiang

Tseng

et al. 2016

IJMS

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The CRISPR/Cas9 Genome-editing system has revealed promising potential for generating gene mutation, deletion, and correction in human cells. Application of this powerful tool in Fabry disease (FD), however, still needs to be explored. Enzyme replacement therapy (ERT), a regular administration of recombinant human α Gal A (rhα-GLA), is a currently available and effective treatment to clear the accumulated Gb3 in FD patients. However, the short half-life of rhα-GLA in human body limits its application. Moreover, lack of an appropriate in vitro disease model restricted the high-throughput screening of drugs for improving ERT efficacy. Therefore, it is worth establishing a large-expanded in vitro FD model for screening potential candidates, which can enhance and prolong ERT potency. Using CRISPR/Cas9-mediated gene knockout of GLA in HEK-293T cells, we generated GLA-null cells to investigate rhα-GLA cellular pharmacokinetics. The half-life of administrated rhα-GLA was around 24 h in GLA-null cells; co-administration of proteasome inhibitor MG132 and rhα-GLA significantly restored the GLA enzyme activity by two-fold compared with rhα-GLA alone. Furthermore, co-treatment of rhα-GLA/MG132 in patient-derived fibroblasts increased Gb3 clearance by 30%, compared with rhα-GLA treatment alone. Collectively, the CRISPR/Cas9-mediated GLA-knockout HEK-293T cells provide an in vitro FD model for evaluating the intracellular pharmacokinetics of the rhα-GLA as well as for screening candidates to prolong rhα-GLA potency. Using this model, we demonstrated that MG132 prolongs rhα-GLA half-life and enhanced Gb3 clearance, shedding light on the direction of enhancing ERT efficacy in FD treatment.

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Section: Discussionmentioning

confidence: 97%

Using CRISPR/Cas9-Mediated GLA Gene Knockout as an In Vitro Drug Screening Model for Fabry Disease

Song¹,

Chiang

Tseng

et al. 2016

IJMS

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“…Moreover, the tools of precise genome editing with engineered nucleases, such as the zinc finger nucleases (zfns), the transcription activator-like effecter nucleases (talens) and, more recently, the Clustered Regularly Interspaced Short Palindromic Repeats (crispr) associated Cas9 technology (Gaj, Gersbach and Barbas, 2013;Kim, 2016;Komor, Badran and Liu, 2017) have opened up tremendous opportunities for the development of cell lines, especially those of human origin (Tobita, Guzman-Lepe and de L'Hortet, 2015). crispr/Cas9 technology was reported for genome editing in hiPSCs (Flaherty and Brennand, 2015;Li et al, 2014;Seah et al, 2015;Suzuki et al, 2014). Another study reported on the simultaneous reprogramming and gene correction of patient fibroblasts (Howden et al, 2015).…”

Section: Toxicology In the 21st Centurymentioning

confidence: 99%

“…Image courtesy of the Wyss Institute, Harvard University .8 Modern cell reprogramming and gene editing tools, allowing modifications of patient-specific iPSCs for use in disease research, toxicology, and screening, in addition to the possibility of cell therapy. Image taken fromSeah et al (2015)…”

mentioning

confidence: 99%

The Changing Paradigm in Preclinical Toxicology: in vitro and in silico Methods in Liver Toxicity Evaluations

Noor¹

2019

Animal Experimentation: Working Towards a Paradigm Change

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“…By contrast, in HDR, a sequence of DNA that is partially homologous (typically the undamaged homologous chromosome) serves as a template for repair (Figure 1). Alternatively, if a DSB occurs in the presence of an exogenously introduced homologous repair template, the addition of specific DNA sequences can occur [51] (Figure 1). …”

Section: Endogenous Repair Of Double-stranded Dna Breaks (Dsb)mentioning

confidence: 99%

May I Cut in? Gene Editing Approaches in Human Induced Pluripotent Stem Cells

Brookhouser

Raman

Potts

et al. 2017

Cells

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In the decade since Yamanaka and colleagues described methods to reprogram somatic cells into a pluripotent state, human induced pluripotent stem cells (hiPSCs) have demonstrated tremendous promise in numerous disease modeling, drug discovery, and regenerative medicine applications. More recently, the development and refinement of advanced gene transduction and editing technologies have further accelerated the potential of hiPSCs. In this review, we discuss the various gene editing technologies that are being implemented with hiPSCs. Specifically, we describe the emergence of technologies including zinc-finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN), and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 that can be used to edit the genome at precise locations, and discuss the strengths and weaknesses of each of these technologies. In addition, we present the current applications of these technologies in elucidating the mechanisms of human development and disease, developing novel and effective therapeutic molecules, and engineering cell-based therapies. Finally, we discuss the emerging technological advances in targeted gene editing methods.

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Induced Pluripotency and Gene Editing in Disease Modelling: Perspectives and Challenges

Cited by 19 publications

References 148 publications

Using CRISPR/Cas9-Mediated GLA Gene Knockout as an In Vitro Drug Screening Model for Fabry Disease

Using CRISPR/Cas9-Mediated GLA Gene Knockout as an In Vitro Drug Screening Model for Fabry Disease

The Changing Paradigm in Preclinical Toxicology: in vitro and in silico Methods in Liver Toxicity Evaluations

May I Cut in? Gene Editing Approaches in Human Induced Pluripotent Stem Cells

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