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Genetic Engineering Publications - GEG Tech top picks
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BigField GEG Tech's insight:
In this study, the authors successfully produced double point mutations in rice acetolactate synthase gene (OsALS) and generated herbicide resistant rice lines by using TALENs and donor DNA carrying the desired mutations. The HR-mediated gene edits were heritable to the progeny of T1 generation. The edited T1 plants were as morphologically normal as the control plants while displayed strong herbicide resistance. The results demonstrate the feasibility of TALEN-mediated genome editing in rice and provide useful information for further genome editing by other nuclease-based genome editing platforms. 
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BigField GEG Tech's insight:
In this study, the authors show that a site specific double strand break (DSB) generated both in the genome and the donor plasmid using the CRISPR-Cas9 system can be efficiently used to target ∼5 kb plasmids into mammalian genomes via nonhomologous end joining (NHEJ). They were able to achieve efficiencies of up to 0.17% in HEK293 cells and 0.45% in CHO cells.
Dominique Blanchard's comment,
May 13, 2015 8:46 AM
I'm going to read this paper because It will greatly impact owners of homologous recombination related patents.
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BigField GEG Tech's insight:
The authors targeted DNA ligase IV, a key enzyme in the NHEJ pathway, using the inhibitor Scr7 to promote HDR at the expense of NHEJ. Scr7 treatment increased the efficiency of HDR-mediated genome editing, using Cas9 in mammalian cell lines and in mice for all four genes examined, up to 19-fold.
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PubMed comprises more than 23 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.
BigField GEG Tech's insight:
CRISPR to modify HPSC by homologous recombination, and one use of more for this technology!
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BigField GEG Tech's insight:
In this work, the scientists compre TALEN and CRISPR to target two loci within the EGFP gene. They find that the CRISPR system induced targeted genomic deletion more efficiently and precisely than TALENs. However, TALENs stimulated homology directed repair more efficiently than CRISPR system and caused fewer targeted genomic deletions. These data suggest that the choice of genome editing tool should be determined by the desired genome editing outcome. Such a rational approach is likely to benefit research outputs for groups working in fields as diverse as modification of cell lines, to animal models for disease studies, or gene therapy strategies.
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BigField GEG Tech's insight:
In this study, the scientists report the use of CRISPR/Cas9 technology to target a specific genomic region in human pluripotent stem cells. They show that CRISPR/Cas9 can be used to disrupt a gene by introducing frameshift mutations to gene coding region; to knock in specific sequences, to targeted genomic locus via homology directed repair and to induce large genomic deletion through dual-guide multiplex.
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BigField GEG Tech's insight:
The authors demonstrated that suppressed the NHEJ key molecules such as KU70 or DNA ligase IV promotes the efficiency of HDR 4–5-fold. When co-expressed with the Cas9 system, E1B55K and E4orf6 improved the efficiency of HDR up to eightfold and essentially abolished NHEJ activity in both human and mouse cell lines.
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BigField GEG Tech's insight:
The authors demonstrated the feasibility to genetically modify the ESCs of Rhesus monkey via CRISPR/Cas9 system for cell tracing.
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Genome editing of human ES/iPS cells has been limited by technical difficulties that result in a low efficiency of homologous recombination (HR) in human ES/iPS cells.
In this work, the authors demonstrated that RAD51 overexpression and valproic acid treatment enhanced biallelic-targeting efficiency in human ES/iPS cells regardless of the transcriptional activity of the targeted locus. Their findings would facilitate genome editing study using human ES/iPS cells.