Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ

Background: Although CRISPR/Cas enables one-step gene cassette knock-in, assembling targeting vectors containing long homology arms is a laborious process for high-throughput knock-in. We recently developed the CRISPR/Cas-based precise integration into the target chromosome (PITCh) system for a gene cassette knock-in without long homology arms mediated by microhomology-mediated end-joining. Results: Here, we identified exonuclease 1 (Exo1) as an enhancer for PITCh in human cells. By combining the Exo1 and PITCh-directed donor vectors, we achieved convenient one-step knock-in of gene cassettes and floxed allele both in human cells and mouse zygotes. Conclusions: Our results provide a technical platform for high-throughput knock-in

A Global Forum on Ultramafic Ecosystems: From Ultramafic Ecology to Rehabilitation of Degraded Environments

The 9th International Conference on Serpentine Ecology (ICSE) was held in Tirana and Pogradec (Albania) from June 5 to 9, 2017. More than 100 delegates from 29 countries around the world gathered to present their research on recent advances in: (i) ultramafic soils, (ii) biogeochemistry, (iii) diversity of ultramafic flora, microflora and fauna, (iv) ecophysiology of ultramafic-adapted organisms, (v) interactions between ultramafic organisms and their ecology, (vi) nature rehabilitation of degraded ultramafic environments (resulting from mining activities), and (vii) the production of bio-based metals through agromining technology. Additionally, the ICSE featured the first symposium on ultramafic aquatic ecology and ecotoxicology. Albania has one of the most diverse ultramafic floras in Europe. During the conference delegates visited some of the most emblematic ultramafic sites in Albania as well as the first agromining field trial in Europe. Here, we present the major topics and provide some highlights of the 25 contributions in this Special Issue (Vol. 33 no.3 and 4)

Oral glucosylceramide reduces 2,4-dinitrofluorobenzene induced inflammatory response in mice by reducing TNF-alpha levels and leukocyte infiltration.

Sphingolipids are constituents of cellular membranes and play important roles as second messengers mediating cell functions. As significant components in foods, sphingolipids have been proven to be critical for human health. Moreover, diverse metabolic intermediates of sphingolipids are known to play key roles both in proinflammatory and in anti-inflammatory effects. However, the effect of dietary sphingolipids on inflammation is a complicated field that needs to be further assessed. Our study evaluated the effects of orally administered maize glucosylceramide (GluCer), one of the most conventional dietary sphingolipids, on inflammation using the 2, 4-dinitro-1-fluorobenzene-treated BALB/c murine model. Oral administration of GluCer inhibited ear swelling and leukocyte infiltration to the inflammatory site, suggesting that dietary GluCer has anti-inflammatory properties. ELISA analyses revealed that oral administration of GluCer for 6 days had not modified the Th1/Th2 balance, but significantly down-regulated the activation of TNF-α at the inflammatory site. Based on these results, the down-regulation of TNF-α by dietary GluCer may suppress vascular permeability and reduce the migration of inflammatory cells. Our findings increase understanding of the actions of dietary sphingolipids on the balance of the immune response

Correction to: A global forum on ultramafic ecosystems: from ultramafic ecology to rehabilitation of degraded environments

The affiliations of the coauthors, Antony van der Ent and Shota Sakaguchi have been incorrectly published in the original publication of the article. The correct affiliations are provided in this correction

Additional file 1: of Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ

The following additional data are available with the online version of this paper. Additional data file 1 contains the figures including IDA analysis of PITCh-donor, PCR screenings of mice, sequencing of non-knock-in Actb alleles, FACS and LSM analysis of human cells, Exo1 western blotting, Exo1 toxicity analysis, off-target analysis in mice, germline transmission, linear PCR donor injection, and sequence alignments of hACTB and its off-target sites, and tables of these results and a list of the oligo DNAs and RNAs used in this study. (DOCX 14447 kb