Genetic Engineering Publications - GEG Tech top picks

In this study, the scientists show that the in vitro application of an HDR enhancer, RS-1, increases the knock-in efficiency by two- to five-fold at different loci, whereas NHEJ inhibitor SCR7 has minimal effects. We then apply RS-1 for animal production and have achieved multifold improvement on the knock-in rates as well. This work presents tools to nuclease-mediated knock-in animal production, and sheds light on improving gene-targeting efficiencies on pluripotent stem cells.

Programmable nucleases, such aZFNs, TALENs,  or CRISPR-Cas9 have the potential for applications in the clinical setting to treat genetic diseases or prevent infectious diseases. However, because the accuracy of DNA recognition by these nucleases is not always perfect, off-target mutagenesis may result in undesirable adverse events. In this review, the scientists provide an overview of available nuclease designing platforms, nuclease engineering approaches to minimize off-target activity, and methods to evaluate both on- and off-target cleavage of CRISPR-Cas9.

www.geg-tech.com/Vectors

Efficient genome-editing tools are needed to correct patient-derived stem cells or model human disease. Hatada et al. report that low (0.4 Gy) doses of g-ray or X-ray radiation can increase recombination efficiency in cells by more than 30-fold when used in combination with zinc finger nucleases, transcription activator.

www.geg-tech.com

This review gives a short primer on gene editing followed by some of the foundational work in gene editing and subsequently a discussion of programmable nucleases leading to a description of Zinc Finger Nuclease, TALENs and CRISPRs.

www.geg-tech.com/Vectors

The authors review the fast developing technology of targeted genome engineering using site specific programmable nucleases zinc finger nucleases (ZFNs), transcription activator like nucleases (TALENs) and cluster regulatory interspaced short palindromic repeat/CRISPR associated proteins (CRISPR/Cas) based RNA-guided DNA endonucleases (RGENs) and their different characteristics including pros and cons of genome modifications by these nucleases. They have further discussed different types of delivery methods to induce gene editing, novel development in genetic engineering other than nucleases and future prospects.

www.geg-tech.com

A major complication with genome editing toolss is the binding of the nuclease to unintended genomic sites that share sequence homology with the on-target site. Here the authors reviewed the significant progress has been made recently to boost the nuclease targeting specificity by protein engineering to modify the structure of the nuclease and alter the interaction with its genomic target.

Here, the authors review the applications of programmable nucleases in biology and medicine with a focus on CRISPR system. They discuss about the impact of the unprecedented ability to modify cells offer by CRISPR to the scientists and about the consequence of the off targets.

www.geg-tech.com/Vectors

In this study, the author compare ZFN and TALEN to target the insertion of a myelo-specific gp91phox cassette to AAVS1. They observe that TALENs were more efficient than ZFNs in generating correctly targeted iPSC colonies, but all corrected iPSC clones showed no signs of mutations at the top-ten predicted off-target sites of both nucleases. These  data open the way to generate autologous, functionally corrected granulocytes with the TALEN system.

www.geg-tech.com/Vectors

“Which technology should I use in my experiment?” This review offers a practical resource to compare and contrast these technologies, guiding the investigator when and where to use this fantastic array of powerful tools.

www.geg-tech.com/Vectors

The authors used zinc finger nucleases (ZFNs) directed against the gene encoding human PD-1 (PDCD-1) to gene-edit melanoma Tumor Infiltrating Lymphocyte (TIL). They show that their clinical scale TIL production process yielded efficient modification of the PD-1 gene locus, with an average modification frequency of 74.8% of the alleles in a bulk TIL population, which resulted in a 76% reduction in PD-1 surface-expression.

www.geg-tech.com/Vectors

The authors describe a protocol for the targeted integration of a doxycycline-inducible transgene expression system in a safe harbor site in iPSCs. This gene targeting strategy uses zinc finger nucleases (ZFNs) to enhance homologous recombination at the AAVS. This protocol could be also compatible with the use of TALEN or CRISPR.

www.geg-tech.com