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BigField GEG Tech's insight:
The authors discuss experimental considerations, limitations and critical aspects which will guide the investigator for successful implementation of the genome editing technology in human PSCs using designer nucleases. 
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BigField GEG Tech's insight:
The authors provide a detailed protocol for design and construction of TALEN and CRISPR vectors, testing of their nuclease activity, and delivery of TALEN or CRISPR vectors into hPSCs. Moreover, they describe the identification of edited hPSC clones without antibiotic selection, including their clonal selection, genotyping, and expansion for downstream applications.
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BigField GEG Tech's insight:
The authors show that activation of the TGFβ pathway in hPSC-derived chondrogenic progenitors promotes the efficient development of articular chondrocytes that can form stable cartilage tissue in vitro and in vivo. In contrast, chondrocytes specified by BMP4 signaling display characteristics of hypertrophy and give rise to cartilage tissues that initiate the endochondral ossification process in vivo.
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BigField GEG Tech's insight:
The scientists used cellular reprogramming and genome engineering to functionally dissect the loss of chromosome 7q, a somatic cytogenetic abnormality present in myelodysplastic syndromes. They used a phenotype-rescue screen to identify candidate haploinsufficient genes that might mediate the del(7q)- hematopoietic defect. Their approach highlights the utility of human iPSCs both for functional mapping of disease-associated large-scale chromosomal deletions and for discovery of haploinsufficient genes.
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BigField GEG Tech's insight:
In this Report, The scientists present an efficient method for seamless correction of pF508del mutation in patient-specific induced pluripotent stem cells by gene edited homologous recombination. Gene edition has been performed by transcription activator-like effector nucleases (TALEN) and a homologous recombination donor vector which contains a PiggyBac transposon-based double selectable marker cassette.
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BigField GEG Tech's insight:
The authors describe methods for rapid synthesis of gRNA and for delivery of Cas9 protein/gRNA ribonucleoprotein complexes (Cas9 RNPs) into a variety of mammalian cells through liposome-mediated transfection or electroporation. Using these methods, they report nuclease-mediated indel rates of up to 94% in Jurkat T cells and 87% in induced pluripotent stem cells (iPSC) for a single target.
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BigField GEG Tech's insight:
To overcome the problem of undesired mutations, the scientists designed strategies to physically destroy or separate CRISPR target sites at the targeted allele and developed a bioinformatic pipeline to identify and eliminate clones harboring deleterious indels at the other allele. This two-pronged approach enables the reliable generation of knockin hPSC reporter cell lines free of unwanted mutations at the targeted locus.
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The authors discuss experimental considerations, limitations and critical aspects which will guide the investigator for successful implementation of the genome editing technology in human PSCs using designer nucleases.