Genetic Engineering Publications - GEG Tech top picks

Forward genetic screens using Sleeping Beauty (SB) mobilized T2/Onc transposons have been used to identify Common Insertion Sites (CIS) associated with tumor formation. Here the authors  developed an automated method to systematically identify T2/Onc-genome RNA fusion sequences in RNA-seq data. RNA fusion based CIS were identified corresponding to both DNA based CIS (Cdkn2a, Mycl1, Nf2, Pten, Sema6d and Rere) and additional regions strongly associated with cancer that were not observed by LM-PCR (Myc, Akt1, Pth, Csf1r, Fgfr2, Wisp1, Map3k5 and Map4k3). These methods independently identify CIS regions, and also point to cancer-associated genes like Braf. They anticipate RNA-seq analyses of tumors from forward genetic screens will become an efficient tool to identify causal events.

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Recently, a simple and inexpensive liver-specific transgenic approach was developed, in which a hydrodynamics-based transfection (HT) method was coupled with the Sleeping Beauty transposase system. Various HT models in which different oncogenic pathways are activated and/or tumor-suppressing pathways inactivated have been developed in recent years. The applicability of HT models in liver cancer research is expected to broaden and ultimately elucidate the cooperation between oncogenic signaling pathways and aid in designing molecular therapy to target altered pathways.

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In this review, diseases affecting the blood system, the connective tissue, the immune system, the metabolism, and the nervous system and their treatment utilizing the SB transposase system will be discussed. Moreover, advantages and disadvantages of SB transposase-based gene therapeutic approaches will be mentioned.

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The authors describe a method for the derivation of bovine iPSCs employing Sleeping Beauty and piggyBac transposon systems encoding different combinations of reprogramming factors, each separated by self-cleaving peptide sequences and driven by the chimeric CAGGS promoter. 

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This review provides a state-of-the-art overview of preclinical studies for treatment of rare genetic diseases based on the SB transposase system for stable correction of the genetic defect. Diseases affecting the blood system, the connective tissue, the immune system, the metabolism, and the nervous system and their treatment utilizing the SB transposase system are discussed.

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In this study, the authors used Sleeping beauty to mediate transposition of hematopoietic progenitors with a transposon encoding a L22Y variant dihydrofolate reductase (DHFR) fused to GFP. Culture of nucleofected cells in medium under selective conditions increased the frequency of GFP+ drug-resistant hematopoietic colony forming cells by 2-fold. 

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In this review, the author discuss the basic structure of screens using sleeping beauty to identify numerous cancer drivers in a wide variety of tumor types.

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The authors report their improvements made to the existing SB vector system and present two new vector types for robust constitutive or inducible expression of any gene of interest.

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