Study on chromatin regulation patterns of expression vectors in the PhiC31 integration site.

The PhiC31 integration system allows for targeted and efficient transgene integration and expression by recognizing pseudo attP sites in mammalian cells and integrating the exogenous genes into the open chromatin regions of active chromatin. In order to investigate the regulatory patterns of efficient gene expression in the open chromatin region of PhiC31 integration, this study utilized Ubiquitous Chromatin Opening Element (UCOE) and activating RNA (saRNA) to modulate the chromatin structure in the promoter region of the PhiC31 integration vector. The study analysed the effects of DNA methylation and nucleosome occupancy changes in the integrated promoter on gene expression levels. The results showed that for the OCT4 promoter with moderate CG density, DNA methylation had a smaller impact on expression compared to changes in nucleosome positioning near the transcription start site, which was crucial for enhancing downstream gene expression. On the other hand, for the SOX2 promoter with high CG density, increased methylation in the CpG island upstream of the transcription start site played a key role in affecting high expression, but the positioning and clustering of nucleosomes also had an important influence. In conclusion, analysing the DNA methylation patterns, nucleosome positioning, and quantity distribution of different promoters can determine whether the PhiC31 integration site possesses the potential to further enhance expression or overcome transgene silencing effects by utilizing chromatin regulatory elements.

A systematic review of microplastics in the environment: Sampling, separation, characterization and coexistence mechanisms with pollutants.

Microplastics (<5 mm) (MPs) are widely distributed throughout the world, and their accumulation and migration in the environment have caused health and safety concerns. Currently, most of the reviewed literatures mainly focus on the distribution in various environmental media, adsorption mechanisms with different pollutants, and characterization of MPs. Therefore, the present review mainly highlights the characterization techniques of MPs and the underlying mechanisms of their combination with conventional coexisting substances (heavy metals, organic pollutants, and nutrients). We observed that massive MP pollution has been found in many areas, especially in Africa, Asia, India, South Africa, North America and Europe. The separation methods of MPs in different environmental media are basically similar, including sampling, pre-treatment, flotation, filtration and digestion. The combination of multiple characterization technologies can more precisely identify the shape, abundance, colour, and particle size of MPs. Notably, although recent reports have confirmed that MPs can act as carriers of heavy metals and carry them into organisms to cause harm, MPs have different adsorption and desorption characteristics for various heavy metals. The adsorption capacity of organic pollutants onto MPs is closely related to their hydrophobicity, specific surface area and functional group characteristics. The relative abundance of MPs in sediments and lakes had a significantly positive correlation with the mass concentration of total nitrogen in lake water, but this finding still needs to be further verified. Based on current research, we suggest that future MP research should focus on characterization technology, environmental migration, ecological effects, health risks and degradation methods.

[Effect of interactions of chromatin regulatory elements with different promoters on the regulation of gene expression].

The effect of altering the promoter region of ubiquitous chromatin-opening element (UCOE) and matrix attachment region (MAR) on stable and efficient expression of genes was investigated. Four different promoters were tested, namely, oct4 containing an enhancer region, sox2 having a CpG island, nanog having no regulatory elements, and CMV containing a CpG island and an enhancer region. Eight reporter plasmids were constructed: pOCT4-UCOE, pOCT4-MAR, pSOX2-UCOE, pSOX2-MAR, pNANOG-UCOE, pNANOG-MAR, pCMV-UCOE, and pCMV-MAR. Stable and efficient expression was observed when UCOE combined with the oct4 promoter, whereas the sox2 was the best promoter suited for MAR. Comparison of the stable clones of oct4-UCOE and sox2-MAR showed that UCOE-regulated expression is more stable and efficient than MAR-regulated expression. When CpG island-containing promoter is linked with UCOE, stable and efficient expression could be observed. These data suggest that an enhancer region in the promoter leads to high, yet unstable expression when combined with UCOE, whereas CpG islands stabilize expression. In conclusion, UCOE and MAR interact with regulatory elements on the promoter by altering the chromatin open state and chromatin loop to regulate gene expression.