Find and replace: editing human genome in pluripotent stem cells

Pan, Huize; Zhang, Weiqi; Zhang, Weizhou; Liu, Guanghui

doi:10.1007/s13238-011-1132-0

Cited by 19 publications

(15 citation statements)

References 54 publications

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“…Targeted gene editing technologies have recently been successfully applied in iPSCs due to their clonal expansion capability [13][14][15][16]. However, current methodologies of direct cell lineage conversion toward terminally committed cells would not be able to provide desired cell types for gene targeting.…”

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confidence: 99%

Induced neural stem cells: a new tool for studying neural development and neurological disorders

Liu

Suzuki

et al. 2012

Cell Res

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confidence: 99%

Induced neural stem cells: a new tool for studying neural development and neurological disorders

Liu

Suzuki

et al. 2012

Cell Res

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“…However, the overall efficiency of obtaining gene-modified cells with this plasmid-based approach is often low. 29,42 A minicircle vector is a new generation of DNA vectors that lack extraneous bacterial sequences encoding antibiotic resistance genes and a bacterial origin of replication. 43,44 Several lines of evidence suggest that the minicircle possesses merits such as robust transgene expression, 43,45 high efficiency in gene transfer [46][47][48] and biosafety, 49 all of which indicate that the minicircle could be an efficient and safe delivery system for programmable nucleases.…”

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confidence: 99%

Enhanced gene disruption by programmable nucleases delivered by a minicircle vector

et al. 2014

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Targeted genetic modification using programmable nucleases such as zinc finger nucleases (ZFNs) and transcription activatorlike effector nucleases (TALENs) is of great value in biomedical research, medicine and biotechnology. Minicircle vectors, which lack extraneous bacterial sequences, have several advantages over conventional plasmids for transgene delivery. Here, for the first time, we delivered programmable nucleases into human cells using transient transfection of a minicircle vector and compared the results with those obtained using a conventional plasmid. Surrogate reporter assays and T7 endonuclease analyses revealed that cells in the minicircle vector group displayed significantly higher mutation frequencies at the target sites than those in the conventional plasmid group. Quantitative PCR and reverse transcription-PCR showed higher vector copy number and programmable nuclease transcript levels, respectively, in 293T cells after minicircle versus conventional plasmid vector transfection. In addition, tryphan blue staining and flow cytometry after annexin V and propidium iodide staining showed that cell viability was also significantly higher in the minicircle group than in the conventional plasmid group. Taken together, our results show that gene disruption using minicircle vector-mediated delivery of ZFNs and TALENs is a more efficient, safer and less toxic method than using a conventional plasmid, and indicate that the minicircle vector could serve as an advanced delivery method for programmable nucleases.Gene Therapy (2014) 21, 921-930;

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“…Gene Editing refers to the process of making targeted modifications to the genome, such as the deletion, insertion or replacement of targeted DNA sequences (Pan et al, 2011). Gene editing technologies fundamentally improve our approaches to biological basic research, gene therapy, genetic improvement, etc.…”

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confidence: 99%