The purpose of this study was to develop an improved zona-free method of goat somatic cell nuclear transfer (SCNT) that has both ease of operation and efficiency. The main steps involved were: (1) optimization of in vitro oocyte maturation, (2) parthenogenetic activation of zona-free oocytes, (3) SCNT of zona-free anaphase II-telophase II (AII-TII) oocytes that subverted the need for long term UV-exposure of the oocytes, and (4) in vitro culture of groups of cloned embryos in wells in a highly efficient continuous serum-free embryo medium to the blastocyst stage before transfer to the recipients. Percentages of transgenic blastocyst production were 22.3 and 33.1% for adult and fetal cell lines, respectively. After transfer of cloned and transgenic blastocysts, 28.6 and 36.4% of the recipients were confirmed pregnant and 75 and 33.3% of the pregnancies resulted in the delivery of viable offspring, respectively. To our knowledge, this is the first report of successful live and survived birth of clo...

http://royanaward.com/files12/s1-ln8561907-1207415488-1939656818Hwf-1293905860IdV-18006427398561907PDF_HI0001.pdf

Development of an optimized zona-free method of somatic cell nuclear transfer in the goat.

In mammals, their proper development during the early cleavage stages strongly relies on the gene products newly transcribed by zygotic genome activation (ZGA). Long noncoding RNAs (lncRNAs) have been characterized as key regulators of the ZGA process in mice and human. However, the ZGA stage has not yet been identified and epigenetic regulations of the ZGA process remain largely unknown in goats. Here, we show that ZGA occurred at the 8-cell stage in goats. During ZGA, trimethylation of H3K9 was dynamically changed but maintained strong staining in development arrested embryos. Using single-cell RNA sequencing, we identified 800 mRNAs and 250 lncRNAs as candidates of key molecules in goat preimplantation embryos. These mRNAs and lncRNAs were differentially expressed from 4- to 8-cell stage embryos and were strongly enriched in terms of retinoic acid receptor signaling pathway as well as signaling pathway regulating pluripotency of stem cells. In particular, we found that microinjection of siRNA against lnc_137 caused development arrest. Our results are consistent with the notion that lncRNAs play vital roles during ZGA, and the data presented here provide an excellent source for further ZGA lncRNA studies.

Long noncoding RNAs exchange during zygotic genome activation in goat.

It has been 30 years since the first transgenic mouse was generated and 26 years since the first example of transferring the technology to livestock was published. While there was tremendous optimism in those initial years, with most convinced that genetically modified animals would play a significant role in agricultural production, that has not come to be. So at first sight one could conclude that this technology has, to a large extent, failed. On the contrary, it is believed that it has succeeded beyond our original expectations, and we are now at what is perhaps the most exciting time in the development and implementation of these technologies. The original goals, however, have drastically changed and it is now biomedical applications that are playing a central role in pushing both technical and scientific developments. The combination of advances in somatic cell nuclear transfer, the development of induced pluripotent stem cells and the completion of the sequencing of most livestock genomes ensures a bright and exciting future for this field, not only in livestock but also in companion animal species.

Perspectives on transgenic livestock in agriculture and biomedicine: an update

Comparative studies on ZIF-8 and SiO₂ nanoparticles as carrier for immobilized β-glucosidase

Background
Somatic cell nuclear transfer (SCNT) is a promising technique to produce transgenic cloned mammalian, including transgenic goats which may produce Human Lactoferrin (hLF). However, success percentage of SCNT is low, because of gestational and neonatal failure of transgenic embryos. According to the studies on cattle and mice, DNA methylation of some imprinted genes, which plays a vital role in the reprogramming of embryo in NT maybe an underlying mechanism.

/pdf/generation-of-five-human-lactoferrin-transgenic-cloned-goats-1u6uy2sick.pdf

Generation of Five Human Lactoferrin Transgenic Cloned Goats Using Fibroblast Cells and Their Methylation Status of Putative Differential Methylation Regions of IGF2R and H19 Imprinted Genes

Production of dairy goat embryos, by nuclear transfer, transgenic for human acid β-glucosidase

TGF-β1 controls porcine granulosa cell states: A miRNA-mRNA network view.

miR-130a/TGF-β1 axis is involved in sow fertility by controlling granulosa cell apoptosis.

Transforming growth factor-β1 (TGF-β1) is essential for ovarian function and female fertility in mammals. Herein, we identified three completely linked variants, including two known variants referred to as c.1583A > G and c.1587A > G and the novel variant c.2074A > C in the porcine TGF-β1 3'-UTR. An important role of these variants in Yorkshire sow fertility was revealed. Variants c.1583A > G and c.1587A > G were located at the miRNA response element (MRE) of miR-2337 and affected miR-2337 regulation of TGF-β1 3'-UTR activity. Interestingly, miR-2337 induces, not reduces the transcription and production of TGF-β1 in granulosa cells (GCs). Mechanistically, miR-2337 enhances TGF-β1 promoter activity via the MRE motif in the core promoter region and alters histone modifications, including H3K4me2, H3K4me3, H3K9me2, and H3K9ac. In addition, miR-2337 controls TGF-β1-mediated activity of the TGF-β signaling pathway and GC apoptosis. Taken together, our findings identify miR-2337 as an endogenous small activating RNA (saRNA) of TGF-β1 in GCs, while miR-2337 is identified as a small activator of the TGF-β signaling pathway which is expected to be a new target for rescuing GC apoptosis and treating low fertility.

/pdf/mir-2337-induces-tgf-b1-production-in-granulosa-cells-by-155rk37cxm.pdf

miR-2337 induces TGF-β1 production in granulosa cells by acting as an endogenous small activating RNA.

Microservices architecture has become the latest trend in building modern applications due to its flexibility, scalability, and agility. However, due to the complex interdependencies between microservices, an anomaly in any one service in a microservice system has the potential to propagate along service dependencies and affect multiple services. Therefore, accurate and efficient root cause localization is a significant challenge for current microservice system operation and maintenance. Focusing on this challenge and leveraging the dynamically constructed service call graph, we propose MicroEGRCL, a root cause localization approach based on graph neural networks with an attention mechanism that includes edge feature enhancement. We conducted an experimental evaluation by injecting various types of service anomalies into two microservice benchmarks running in a Kubernetes cluster. The experimental results demonstrate that MicroEGRCL can achieve an average top1 localization accuracy of 87%, exceeding the state-of-the-art baseline approaches. 

Lingfang Wang

Papers

Production of dairy goat embryos, by nuclear transfer, transgenic for human acid β-glucosidase

TGF-β1 controls porcine granulosa cell states: A miRNA-mRNA network view.

miR-130a/TGF-β1 axis is involved in sow fertility by controlling granulosa cell apoptosis.

miR-2337 induces TGF-β1 production in granulosa cells by acting as an endogenous small activating RNA.

MicroEGRCL: An Edge-Attention-Based Graph Neural Network Approach for Root Cause Localization in Microservice Systems