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Genetic Engineering Publications - GEG Tech top picks
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BigField GEG Tech's insight:
The scientists report the results of subretinal transplantation of human embryonic-stem-cell (hESC)-derived retinal pigment epithelium in four Asian patients: two with dry age-related macular degeneration and two with Stargardt macular dystrophy. They were followed for 1 year. There was no evidence of adverse proliferation, tumorigenicity, ectopic tissue formation, or other serious safety issues related to the transplanted cells. Visual acuity improved 9–19 letters in three patients and remained stable (+1 letter) in one patient. The results confirmed that hESC-derived cells could serve as a potentially safe new source for regenerative medicine. 
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BigField GEG Tech's insight:
The scientists describe the simultaneous transfection of two paired CRISPR sgRNAs-Cas9 plasmids, in mouse embryonic stem cells (mESCs), resulting in the knockout of the selected target gene. Together with a four primer-evaluation system, it poses an efficient way to generate new independent knockout mouse embryonic stem cell lines.
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BigField GEG Tech's insight:
The authors compare the genetic and epigenetic integrity of human nuclear-transfer embryonic stem cell (NT-ESC) lines and isogenic induced pluripotent stem cell (iPSC) lines. As iPSCs, NT-ESCs displayed clone- and gene-specific aberrations in DNA methylation and allele-specific expression of imprinted genes. The occurrence of these genetic and epigenetic defects in both NT-ESCs and iPSCs suggests that they are inherent to reprogramming, regardless of derivation approach.
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BigField GEG Tech's insight:
Here, the authors introduce and optimize a microfluidic membrane deformation method to deliver sgRNA and Cas9 into different cell types and achieve successful genome editing, including for hard-to-transfect lymphoma cells and embryonic stem cells, This method could potentially enable new avenues of biomedical research and gene targeting therapy such as mutation correction of disease genes through combination of the CRISPR-Cas9–mediated knockin system.
Ally Evans's curator insight,
January 4, 2016 3:24 AM
identifying CRISPR/Cas-induced mutations via deep sequencing.
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BigField GEG Tech's insight:
Thanks to the CRISPR/Cas9 system, the authors demonstrate discrete and opposing roles for the lncRNA transcript Haunt and its genomic locus in regulating the HOXA gene cluster during ESC differentiation. Reducing or enhancing Haunt expression, with minimal disruption of the Haunt locus, led to upregulation or downregulation of HOXA genes, respectively. In contrast, increasingly large genomic deletions within the Haunt locus attenuated HOXA activation.
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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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BigField GEG Tech's insight:
The authors describe a protocol to convert human induced pluripotent stem cells or embryonic stem cells into cells similar to cord-blood endothelial colony–forming cells (CB-ECFCs). These CB-ECFC-like cells display a stable endothelial phenotype with high clonal proliferative potential and the capacity to form human vessels in mice and to repair the ischemic mouse retina and limb, and they lack teratoma formation potential.
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Here, the scientists describe the application of an inducible, RNA-guided, nuclease-deficient Cas9-KRAB system to silence target gene expression in human embryonic stem cells, via KRAB repression at the promoter region. This chapter outlines a detailed protocol for generation of a stable human embryonic stem cell line containing both Sp-dCas9-KRAB and sgRNA, followed by inducible expression of Sp-dCas9-KRAB to analyze functional effects of dCas9-KRAB at target loci in human embryonic stem cells.