Promoter Function Research Articles

Mechanism on CMD reaction regulated by supports and promoters of Fe-based catalysts

Methane cracking technology for low-carbon energy conversion to produce hydrogen is a key technical approach in the pursuit of fossil energy decarbonization. Catalytic methane decomposition (CMD) is a process that accomplishes the fundamental removal of carbon from fossil energy without oxygen, producing high-quality carbon-based materials and H2. The functionality of supports and promoters was investigated using inorganic metal oxide support Al2O3, supported active metal Fe, and cocatalyst K. The results show that the Al2O3–Fe catalyst achieves the highest CH4 conversion of 65% at 850 °C, with a gas-phase H2 yield of 1189.72 mmol/gFe within 3 h, and a solid-phase carbon yield of 6.32 g/gFe, significantly higher than at 750 °C. The Al2O3–Fe–K catalyst achieves a methane conversion rate of 66.28% at 850 °C for 1.5 h, exhibiting relatively fast deactivation. The Al2O3–Fe catalyst catalyzes the formation of carbon nanotubes (CNTs) through methane cracking, resulting in mainly 'chain-locked' morphology. The promoter K acts on the surrounding metal clusters to influence the dispersion of active metal. The Al2O3–Fe–K catalyst is more favorable for the production of 'bamboo-shaped' carbon nanotubes, suggesting that K has a more significant impact on shaping the CNTs' products.

Omnichannel as a trend in managing marketing communications of microfinance business in the digital economy

The intense development of the competitive environment requires microfinance organizations to implement highly effective marketing, aimed at achieving commercial goals and realizing promotion functions in the digital environment. Omnichannel marketing becomes relevant in such conditions, emerging as a widespread trend in the contemporary microfinance business. This work reveals the modern trends of omnichannel marketing development: digitalization, the predominance of large commercial structures, and the shift towards a customer–centric approach and omnichannel marketing. Specific tools of omnichannel marketing are highlighted and analysed: email marketing, call centers, chatbots, messengers, websites, and others, as well as some methods of managing loyalty and personalizing interaction with clients are considered. The research concludes that omnichannel marketing not only contributes to the competitiveness of microfinance organizations through improving customer experience and increasing loyalty but also serves as a significant and effective tool for ensuring efficiency and achieving the marketing commercial goals of the microfinance business.

Identification of a sugarcane bacilliform virus promoter that is activated by drought stress in plants

Sugarcane (Saccharum spp.) is an important sugar and biofuel crop in the world. It is frequently subjected to drought stress, thus causing considerable economic losses. Transgenic technology is an effective breeding approach to improve sugarcane tolerance to drought using drought-inducible promoter(s) to activate drought-resistance gene(s). In this study, six different promoters were cloned from sugarcane bacilliform virus (SCBV) genotypes exhibiting high genetic diversity. In β-glucuronidase (GUS) assays, expression of one of these promoters (PSCBV-YZ2060) is similar to the one driven by the CaMV 35S promoter and >90% higher compared to the other cloned promoters and Ubi1. Three SCBV promoters (PSCBV-YZ2060, PSCBV-TX, and PSCBV-CHN2) function as drought-induced promoters in transgenic Arabidopsis plants. In Arabidopsis, GUS activity driven by promoter PSCBV-YZ2060 is also upregulated by abscisic acid (ABA) and is 2.2–5.5-fold higher when compared to the same activity of two plant native promoters (PScRD29A from sugarcane and PAtRD29A from Arabidopsis). Mutation analysis revealed that a putative promoter region 1 (PPR1) and two ABA response elements (ABREs) are required in promoter PSCBV-YZ2060 to confer drought stress response and ABA induction. Yeast one-hybrid and electrophoretic mobility shift assays uncovered that transcription factors ScbZIP72 from sugarcane and AREB1 from Arabidopsis bind with two ABREs of promoter PSCBV-YZ2060. After ABA treatment or drought stress, the expression levels of endogenous ScbZIP72 and heterologous GUS are significantly increased in PSCBV-YZ2060:GUS transgenic sugarcane plants. Consequently, promoter PSCBV-YZ2060 is a possible alternative promoter for genetic engineering of drought-resistant transgenic crops such as sugarcane.

The human MIF polymorphism CATT7 enhances pro-inflammatory macrophage polarization in a clinically relevant model of allergic airway inflammation.

High level expression of the pro-inflammatory cytokine macrophage migration inhibitory factor (MIF) has been associated with severe asthma. The role of MIF and its functional promotor polymorphism in innate immune training is currently unknown. Using novel humanized CATT7 MIF mice, this study is the first to investigate the effect of MIF on bone marrow-derived macrophage (BMDM) memory after house dust mite (HDM) challenge. CATT7 BMDMs demonstrated a significant primed increase in M1 markers following HDM and LPS stimulation, compared to naive mice. This M1 signature was found to be MIF-dependent, as administration of a small molecule MIF inhibitor, SCD-19, blocked the induction of this pro-inflammatory M1-like phenotype in BMDMs from CATT7 mice challenged with HDM. Training naive BMDMs in vitro with HDM for 24 h followed by a rest period and subsequent stimulation with LPS led to significantly increased production of the pro-inflammatory cytokine TNFα in BMDMs from CATT7 mice but not WT mice. Addition of the pan methyltransferase inhibitor MTA before HDM training significantly abrogated this effect in BMDMs from CATT7 mice, suggesting that HDM-induced training is associated with epigenetic remodelling. These findings suggest that trained immunity induced by HDM is under genetic control, playing an important role in asthma patients with the high MIF genotypes (CATT6/7/8).

Will the beneficial properties of plant-growth promoting bacteria be affected by waterlogging predicted in the wake of climate change: A model study

The agricultural sector today is faced with climate change that amplifies extreme climatic events. The use of plant growth promoting bacteria (PGPB) could provide a new approach to increase crop production under these new conditions. In this study, PGPB were isolated from a 5-week pot experiment conducted under greenhouse conditions using cabbage (Brassica oleracea L. var. capitata f. alba) as a model plant. Plants were exposed to either one or two long-term waterlogging events at two different stages of plant development. The results clearly showed that waterlogging leads to changes in the soil microbial community affecting both its overall structure (culturable bacterial fraction) as well as the plant growth promotion (PGP) function provided by the beneficial bacteria to the plant. In general, “the core” of the bacterial community (Bacillus, Pseudomonas, Stenotrophomonas and Peribacillus populations) remained preserved under waterlogged conditions, but shifts in their abundance were observed, with Bacillus, and Pseudomonas proliferating under these conditions. The key to adaptation to waterlogging may lie in “rare” bacterial populations isolated only under certain waterlogging conditions. In our study we identified 11 different genera that can be termed as waterlogging-tolerant. Waterlogging was also shown to induce shifts in the PGP potential, as well as on the structure of a population exhibiting a particular trait, which could eventually lead to either promotion or reduction of plant growth. While the abundances of populations promoting nutrient availability to plants were found to increase after waterlogging, biocontrol protease activity proved to be the trait most sensitive to waterlogging. Enterobacter and Pantoea species were identified as main players in biofilm formation under waterlogging conditions. Waterlogging also appears to trigger the emergence of extremophilic and acidophilic bacteria, aspect important for potential adaptation of soil bacteria to future climate projections, with Bacillus and Pseudomonas showing best potential. Based on all PGP characteristics studied, we selected 11 bacterial isolates belonging to genera Enterobacter, Bacillus, Pseudomonas, Stenotrophomonas, and Pantoea designating them as the most promising bacterial bioinoculants i.e. strains with good potential for future applications in potential waterstress recover of the soil and plants.

Identification and functional analysis of DNA methylation-related enzyme gene family in Eleutherococcus senticosus

Eleutherococcus senticosus, a medicinal and edible plant recognized historically in East Asia, is characterized by a rich composition of saponins and other biologically active ingredients. In this study, 11 cytosine-5 DNA methyltransferases (C5-MTase) and 6 demethylases (dMTase) genes were identified within the genome of E. senticosus. Specifically, two major EsC5-MTases (EsMET1a and EsCMT3b) and two EsdMTases (EsROS1a and EsDME1) were screened through correlation analysis, examining the relationship between their gene expression levels and DNA methylation ratio within E. senticosus genome. Promoter function analysis revealed that the expression of these four prominent genes was regulated by abiotic stresses such as drought and light. In vitro catalytic validation demonstrated the functionality of EsMET1a, EsCMT3b, EsROS1a, and EsDME1 proteins in modifying DNA methylation patterns of key enzyme genes associated with saponin synthesis in E. senticosus. Additionally, transient in vivo overexpression of the four central genes could alter the saponin content by modulating the DNA methylation levels and secondary metabolic processes in E. senticosus. Molecular docking studies further elucidated the possible mechanism of DNA methylation and demethylation catalyzed by these four primary proteins.

Ubiquitin-specific protease 21 promotes tumorigenicity and stemness of colorectal cancer by deubiquitinating and stabilizing ZEB1.

Colorectal cancer (CRC) is one very usual tumor together with higher death rate. Ubiquitin-specific protease 21 (USP21) has been confirmed to take part into the regulation of CRC progression through serving as a facilitator. Interestingly, the promotive function of USP21 has also discovered in the progression of CRC. ZEB1 has illustrated to be modulated by USP7, USP22 and USP51 in cancers. However, the regulatory functions of USP21 on ZEB1 in CRC progression need more investigations. To investigate the relationship between USP21 and ZEB1 in CRC progression. The mRNA and protein expressions were assessed through RT-qPCR, western blot and IHC assay. The interaction between USP21 and ZEB1 was evaluated through Co-IP and GST pull down assays. The cell proliferation was detected through colony formation assay. The cell migration and invasion abilities were determined through Transwell assay. The stemness was tested through sphere formation assay. The tumor growth was evaluated through in vivo mice assay. In this work, USP21 and ZEB1 exhibited higher expression in CRC, and resulted into poor prognosis. Moreover, the interaction between USP21 and ZEB1 was further investigated. It was demonstrated that USP21 contributed to the stability of ZEB1 through modulating ubiquitination level. In addition, USP21 strengthened cell proliferation, migration and stemness through regulating ZEB1. At last, through in vivo assays, it was illustrated that USP21/ZEB1 axis aggravated tumor growth. For the first time, these above findings manifested that USP21 promoted tumorigenicity and stemness of CRC by deubiquitinating and stabilizing ZEB1. This discovery suggested that USP21/ZEB1 axis may provide novel sights for the treatment of CRC.

Viral reprogramming of host transcription initiation.

Viruses are master remodelers of the host cell environment in support of infection and virus production. For example, viruses typically regulate cell gene expression through modulating canonical cell promoter activity. Here, we show that Epstein Barr virus (EBV) replication causes 'de novo' transcription initiation at 29674 new transcription start sites throughout the cell genome. De novo transcription initiation is facilitated in part by the unique properties of the viral pre-initiation complex (vPIC) that binds a TATT[T/A]AA, TATA box-like sequence and activates transcription with minimal support by additional transcription factors. Other de novo promoters are driven by the viral transcription factors, Zta and Rta and are influenced by directional proximity to existing canonical cell promoters, a configuration that fosters transcription through existing promoters and transcriptional interference. These studies reveal a new way that viruses interact with the host transcriptome to inhibit host gene expression and they shed light on primal features driving eukaryotic promoter function.

The study of the factors of the creative industries functioning efficiency

Aim. To investigate the dependence of the effectiveness of the functioning of creative industries on the presence of creative network nodes in regional localization.Objectives. Collection of statistical information on the number and results of the work of organizations belonging to the complex of creative industries; determination of the fullness of the creative industries network in the localization of the regions of the Russian Federation (RF); correlation analysis.Methods. The authors used the methods of statistical data processing, comparative and correlation analysis, general scientific methods of comparison, generalization and concretization.Results. The conducted research confirmed the fact of high connectedness of organizations functioning within one creative network. A high correlation between the performance indicators of the organizations forming the creative product and the presence in the regional location of organizations that perform the functions of promotion and distribution of the creative product has been revealed.Conclusions. The obtained results confirm the high relevance of using the methodology of network approach in the study and development of creative industries. Formation of managerial decisions to stimulate the development of creative economy in the localization of regions should include complex decisions regarding all participants of the creative network.

Effects of Tumor Microenvironment on the Primo Vascular Pattern in the Mouse Model of Metastatic Breast Cancer.

: Tumor survival, promotion, and metastatic functions are regulated by the tumor microenvironment (TME). The primo vascular system (PVS), the third circulatory system in animals, is currently thought to be a highly effective pathway for the spread of cancer cells. : In the present study, we intend to determine the TME effects on the PVS pattern in breast cancer for the first time. : Heterotopic and orthotopic metastatic triple-negative breast cancer (TNBC) mice models were created. After 35 days, the skin was retracted, and a 2 cm skin incision was made up and down from the surface of the tumor tissue. In preparation for PVS staining, the dyes (trypan blue and alamarBlue) were injected throughout the tumor tissues. Under a stereomicroscope, PVS in heterotopic and orthotopic tumors was seen. : According to our data, there are no appreciable variations in PVS patterns and density between heterotopic and orthotopic animal models. Furthermore, alamarBlue is a good option for tumor PVS staining, as demonstrated by our research. : For the first time, our data gave significant new information about the PVS in TNBC. Creating new anti-cancer treatments may be made possible by a better understanding of the biological characteristics of the TME and PVS.

Gene-edited Mtsoc1 triple mutant Medicago plants do not flower.

Optimized flowering time is an important trait that ensures successful plant adaptation and crop productivity. SOC1-like genes encode MADS transcription factors, which are known to play important roles in flowering control in many plants. This includes the best-characterized eudicot model Arabidopsis thaliana (Arabidopsis), where SOC1 promotes flowering and functions as a floral integrator gene integrating signals from different flowering-time regulatory pathways. Medicago truncatula (Medicago) is a temperate reference legume with strong genomic and genetic resources used to study flowering pathways in legumes. Interestingly, despite responding to similar floral-inductive cues of extended cold (vernalization) followed by warm long days (VLD), such as in winter annual Arabidopsis, Medicago lacks FLC and CO which are key regulators of flowering in Arabidopsis. Unlike Arabidopsis with one SOC1 gene, multiple gene duplication events have given rise to three MtSOC1 paralogs within the Medicago genus in legumes: one Fabaceae group A SOC1 gene, MtSOC1a, and two tandemly repeated Fabaceae group B SOC1 genes, MtSOC1b and MtSOC1c. Previously, we showed that MtSOC1a has unique functions in floral promotion in Medicago. The Mtsoc1a Tnt1 retroelement insertion single mutant showed moderately delayed flowering in long- and short-day photoperiods, with and without prior vernalization, compared to the wild-type. In contrast, Mtsoc1b Tnt1 single mutants did not have altered flowering time or flower development, indicating that it was redundant in an otherwise wild-type background. Here, we describe the generation of Mtsoc1a Mtsoc1b Mtsoc1c triple mutant lines using CRISPR-Cas9 gene editing. We studied two independent triple mutant lines that segregated plants that did not flower and were bushy under floral inductive VLD. Genotyping indicated that these non-flowering plants were homozygous for the predicted strong mutant alleles of the three MtSOC1 genes. Gene expression analyses using RNA-seq and RT-qPCR indicated that these plants remained vegetative. Overall, the non-flowering triple mutants were dramatically different from the single Mtsoc1a mutant and the Arabidopsis soc1 mutant; implicating multiple MtSOC1 genes in critical overlapping roles in the transition to flowering in Medicago.

High-resolution RNA tertiary structures in Zika virus stem-loop A for the development of inhibitory small molecules.

Flaviviruses such as Zika (ZIKV) and dengue virus (DENV) are positive-sense RNA viruses belonging to Flaviviridae The flavivirus genome contains a 5' end stem-loop promoter sequence known as stem-loop A (SLA) that is recognized by the flavivirus polymerase NS5 during viral RNA synthesis and 5' guanosine cap methylation. The crystal structures of ZIKV and DENV SLAs show a well-defined fold, consisting of a bottom stem, side loop, and top stem-loop, providing unique interaction sites for small molecule inhibitors to disrupt the promoter function. To facilitate the identification of small molecule binding sites in flavivirus SLA, we determined high-resolution structures of the bottom and top stems of ZIKV SLA, which contain a single U- or G-bulge, respectively. Both bulge nucleotides exhibit multiple orientations, from folded back on the adjacent nucleotide to flipped out of the helix, and are stabilized by stacking or base triple interactions. These structures suggest that even a single unpaired nucleotide can provide flexibility to RNA structures, and its conformation is mainly determined by the stabilizing chemical environment. To facilitate discovery of small molecule inhibitors that interfere with the functions of ZIKV SLA, we screened and identified compounds that bind to the bottom and top stems of ZIKV SLA.

Injectable and Thermosensitive Hydrogel with Platelet-Rich Plasma for Enhanced Biotherapy of Skin Wound Healing.

The rapid and effective healing of skin wounds resulted from severe injuries and full-layer skin defects remains a pressing clinical challenge in contemporary medical practice. The reduction of wound infection and rapid healing is helpful to rebuild and repair skin tissue. Here, a thermosensitive chitosan-based wound dressing hydrogel incorporating β-glycerophosphate (GP), hydroxy propyl cellulose (HPC), graphene oxide (GO), and platelet-rich plasma (PRP) is developed, which exhibits the dual functions of antibacterial properties and repair promotion. GP and HPC enhance the mechanical properties through forming hydrogen bonding connection, while GO produces local heat under near-infrared light, leading to improved blood circulation and skin recovery. Notably, antibacterial properties against Pseudomonas aeruginosa, and control-release of growth factors from PRP are also achieved based on the system. In vitro experiments reveal its biocompatibility, and ability to promote cell proliferation and migration. Animal experiments demonstrate that the epithelial repair and collagen deposition can be promoted during skin wound healing in Sprague Dawley rats. Moreover, a reduction in wound inflammation levels and the improvement of wound microenvironment are observed, collectively fostering effective wound healing. Therefore, the composite hydrogel system incorporated with GO and PRP can be a promising dressing for the treatment of skin wounds.

Transcription activation in Escherichia coli and Salmonella.

Promoter-specific activation of transcript initiation provides an important regulatory device in Escherichia coli and Salmonella. Here, we describe the different mechanisms that operate, focusing on how they have evolved to manage the "housekeeping" bacterial transcription machinery. Some mechanisms involve assisting the bacterial DNA-dependent RNA polymerase or replacing or remodeling one of its subunits. Others are directed to chromosomal DNA, improving promoter function, or relieving repression. We discuss how different activators work together at promoters and how the present complex network of transcription factors evolved.

Can green space exposure enhance the health of rural migrants in Wuhan, China? An exploration of the multidimensional roles of place attachment

Numerous studies have yielded evidence supporting the beneficial health effects of increased green space exposure (GSE), particularly for urban populations. Nevertheless, the specific impacts of GSE on the health of rural migrants, who are regarded as an environmentally vulnerable group, remain unclear. Taking Wuhan, China, as the typical case, this paper examines the nuanced relationship between the multi-facets of GSE and rural migrants’ physical and mental health and explores the mediating roles of multidimensional place attachment (P-AT), including place dependence (P-DE), place identity (P-ID), and place integration (P-IN), based on multisource data. The results are as follows: (1) Different attributes of GSE produced varying effects on rural migrants’ health. Positive correlations emerged between physical health and the availability and accessibility of green space (GS), while mental health exhibited positive associations with visibility and accessibility. (2) There is a complicated landscape in terms of the mediating effects of P-AT. Specifically, P-DE, P-ID, and P-IN acted as mediators between accessibility and the physical and mental health of rural migrants. Concerning availability's influence on physical health, P-ID, and P-IN served as mediators. P-ID mediated the visibility-mental health relationship. Our research underscores the significance of conceptually decomposing the GSE and P-AT in studies on the environment and the health of rural migrants and their diverse social functions in health promotion from GS. We advocate for tailored strategies by local authorities for the migrant population, including maximizing the opportunities of GSE, promoting the restoration and construction of wild GS, and facilitating the sociality of GS to enhance rural migrants’ place sentiment.

Research on the motivation system and path simulation of collaborative agglomeration of Chinese culture and tourism industries based on system dynamics.

In the context of industrial integration, the collaborative agglomeration of the cultural and tourism industries is an important way to promote their integrated development and achieve both industrial transformation and upgrading. This article first analyzes the dynamics of the cultural and tourism industries as a collaborative agglomeration. A system dynamics model is then presented which represents the perspective and reveals the mechanics of this partnership between the two industries. Finally, the authors use this model to simulate the path made possible by their collaborative agglomeration. The results show: (1) From the perspective of industrial policy, the promotion and guidance function of industrial policy elements still needs strengthening in the collaborative development of China's cultural and tourism industries. (2) From the perspective of industrial economy, the promotion function of industrial economic factors still needs improving in the collaborative agglomeration and development of China's cultural and tourism industries. (3) From the perspective of the joint effect of industrial policy and economy, the collaborative effect of industrial policies and economic factors is more conducive to promoting the collaborative agglomeration development of China's cultural and tourism industries. The research in this article can provide theoretical support and policy recommendations for promoting coordinated development of China's cultural and tourism industries and can also provide the experience needed to serve as a reference for the joint development of tourism and culture in other similar regions.

Experimental Studies on the Effects of Glycitinregulated TGF-β Signaling Pathway on the Proliferation, Osteogenic, and Lipogenic Differentiation of BMSCs

Background Glycitin, generated from the seeds of the legume soybean, has an essential function in antioxidant, obesity inhibition, and wound healing promotion. Furthermore, recent research has shown that it has anti-inflammatory and cartilage-protective properties. Objectives This study was designed to investigate the osteogenic effects of glycitin and the transforming growth factor-β (TGF-β) signaling pathway on bone marrow mesenchymal stem cells (BMSCs), as well as their interaction. Materials and Methods We isolated rabbit BMSCs by whole bone marrow adherent method and cultured, identified, and induced differentiation. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was used to determine the proliferation ability of BMSCs with different concentrations of glycitin at different times; reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression levels of osteogenesis-related genes runt-related transcription factor 2 (Runx2), collagen type I (Col-1), and osteocalcin (OC) mRNA; and a kit was used to determine the activities of alkaline phosphatase and triglycerides (TG) in BMSCs. A Western blot was used to detect the expression of TGF-β protein in each group. Results On the fifth day of BMSC primary culture, the morphology of long spindle-shaped cells might be plentiful, dominated by a daisy-like or whirling arrangement. They proliferated rapidly and fused to 70%–80% on the eighth day of culture; the third generation of BMSCs was taken for phenotyping, and the detection results of flow cytometry analysis showed that the surface antigenic markers of BMSCs, CD44, and CD90 were positive, whereas CD34 and CD45 were negative; after osteogenic induction of BMSCs, visible calcified nodules were evident. There was cartilage matrix formation after osteogenic induction; glycitin promoted BMSC proliferation in a time-dependent and dose-dependent manner, and BMSC proliferation increased significantly at 20 µM glycitin for 3–5 days; glycitin increased the expression of Runx2, Col-1, and OC mRNA in BMSCs and increased the activity of alkaline phosphatase (ALP) while decreasing the activity of TG. Meanwhile, 20 µM glycitin dramatically increased the expression of TGF-β. Conclusion These findings imply that glycitin increases BMSCs proliferation and osteogenic differentiation. By regulating the TGF-β signaling pathway, glycitin modulates the differentiation of BMSCs into osteoblasts.

Evidence for novel mechanisms that control cell-cycle entry and cell size.

Entry into the cell cycle in late G1 phase occurs only when sufficient growth has occurred. In budding yeast, a cyclin called Cln3 is thought to link cell-cycle entry to cell growth. Cln3 accumulates during growth in early G1 phase and eventually helps trigger expression of late G1 phase cyclins that drive cell-cycle entry. All current models for cell-cycle entry assume that expression of late G1 phase cyclins is initiated at the transcriptional level. Current models also assume that the sole function of Cln3 in cell-cycle entry is to promote transcription of late G1 phase cyclins, and that Cln3 works solely in G1 phase. Here, we show that cell cycle-dependent expression of the late G1 phase cyclin Cln2 does not require any functions of the CLN2 promoter. Moreover, Cln3 can influence accumulation of Cln2 protein via posttranscriptional mechanisms. Finally, we show that Cln3 has functions in mitosis that strongly influence cell size. Together, these discoveries reveal the existence of surprising new mechanisms that challenge current models for control of cell-cycle entry and cell size.

Ninjurin 2, a Cell Adhesion Molecule and a Target of p53, Modulates Wild-Type p53 in Growth Suppression and Mutant p53 in Growth Promotion.

The nerve injury-induced protein 1 (NINJ1) and NINJ2 constitute a family of homophilic adhesion molecules and are involved in nerve regeneration. Previously, we showed that NINJ1 and p53 are mutually regulated and the NINJ1-p53 loop plays a critical role in p53-dependent tumor suppression. However, the biology of NINJ2 has not been well-explored. By using multiple in vitro cell lines and genetically engineered mouse embryo fibroblasts (MEFs), we showed that NINJ2 is induced by DNA damage in a p53-dependent manner. Moreover, we found that the loss of NINJ2 promotes p53 expression via mRNA translation and leads to growth suppression in wild-type p53-expressing MCF7 and Molt4 cells and premature senescence in MEFs in a wild-type p53-dependent manner. Interestingly, NINJ2 also regulates mutant p53 expression, and the loss of NINJ2 promotes cell growth and migration in mutant p53-expressing MIA-PaCa2 cells. Together, these data indicate that the mutual regulation between NINJ2 and p53 represents a negative feedback loop, and the NINJ2-p53 loop has opposing functions in wild-type p53-dependent growth suppression and mutant p53-dependent growth promotion.

Exploring the Function of Gene Promoter Regulatory Elements Using CRISPR Tools.

Gene promoters serve as pivotal regulators of transcription, orchestrating the initiation, rate, and specificity of gene expression, resulting in cellular diversity found among distinct cell types within multicellular organisms. Identification of the sequence and function of promoters' regulatory elements and their complex interaction with transcription factors, enhancers, silencers, and insulators is fundamental to coordinated transcriptional processes within cells. Identifying these regulatory elements and scrutinizing their functions and interactions through the use of synthetic promoters can pave the way for researchers in various fields ranging from uncovering the origins of diseases associated with promoter mutations to harnessing these regulatory components in biotechnological applications.In this chapter, we describe the manipulation of regulatory elements within promoters, with a specific focus on the use of CRISPR technology on enhancers and silencer elements of the Ovalbumin gene promoter. We explain and discuss processes for the deletion of/interference with regulatory elements within the promoter, employing CRISPR-based approaches. Furthermore, we demonstrate that a CRISPR/Cas-manipulated promoter can activate gene transcription in cell types where it is normally inactive. This confirms that CRISPR technology can be effectively used to engineer synthetic promoters with desired characteristics, such as inducibility, tissue-specificity, or enhanced transcriptional strength. Such an approach provides valuable insights into the mechanisms and dynamics of gene expression, thereby offering new opportunities in the fields of biotechnology and medicine.