In recent years, inducing pluripotent stem cells to differentiate into functional primordial germ (PGCs) in vitro has become an important method of obtaining a large number PGCs. However, the instability and low induction efficiency PGC system restrict application PGCs transgenic animal production, germplasm resource conservation other fields. this study, we successfully established two-step model chicken vitro, which significantly improved formation PGC-like (PGCLCs). To further improve...

Aromatase, a crucial enzyme for estrogen synthesis, plays vital role in gender determination and differentiation. This study aimed to establish an inducible knockout model of the chicken CYP19A1 gene, which encodes aromatase, support control chickens. We selected most efficient sgRNA target site constructed based on Tet-on system. The efficiency reached 80% with 20 μg/mL DOX induction vitro. encapsulation plasmid PEI injection into eggs achieved 45% ovo. qRT-PCR analysis revealed significant...

Insulin is an important component of stem cell cultures; however, its role in the proliferation avian primordial germ cells (PGCs) unknown. The PGCs cultures varies and growth factors signaling pathways necessary to induce chickens are Therefore, we conducted present study investigate effect insulin on survival PGCs. In this study, observed that under culture system, proliferate presence insulin, but do not absence insulin. Furthermore, insulin-lacking media, expression pluripotency genes,...

Myostatin (MSTN) is an important gene involved in the regulation of embryonic muscle cells and adult development; it has a good application prospect transgenic animal production by improving yield muscle. The purpose this study to construct MSTN knockout vector using clustered regularly interspaced short palindromic repeats ( CRISPR)/CRISPR-associated protein 9 Cas9). efficiency was evaluated sheep ear fibroblasts (SEFs) cleavage activity transcription guide RNA gRNA),...

The acetyl CoA acyltransferase 2 (ACAA2) is a key enzyme of the fatty acid oxidation pathway, catalyzing last step mitochondrial beta oxidation, thus playing an important role in metabolism. purpose this study was to investigate effect knocking out ACAA2 on expression genes lipoprteinlipase (LPL), peroxisome proliferator-activated receptor-γ (PPAR-γ), synthase, fat mass and obesity-associated gene, adipocyte acid-binding protein (AP2) precursor adipocytes their differentiation into...

An efficient genome editing approach had been established to construct the stable transgenic cell lines in domestic chicken (Gallus gallus domesticus) at present. Our objectives were investigate gene function differentiation process of embryonic stem cells (ESCs) into spermatogonial cells(SSCs). Three guides RNA (gRNAs) designed knockout Stra8 gene, and efficiency was evaluated using cleavage activity vitro transcription gRNA, Luciferase-SSA assay, T7 endonuclease I assay(T7E1) TA clone...

Abstract The present study established an efficient genome editing approach for the construction of stable transgenic cell lines domestic chicken (Gallus gallus domesticus). Our objectives were to facilitate breeding high-yield, high-quality strains, and investigate gene function in stem cells. Three guide RNA (gRNAs) designed knockout C2EIP gene, efficiency was evaluated DF-1 fibroblasts ESCs using luciferase single-strand annealing (SSA) recombination assay, T7 endonuclease I (T7EI) TA...

The production of germ cells in vitro would open important new avenues for stem biology and human medicine, but the mechanisms cell differentiation are not well understood. chicken, as a great model embryology development, was used this study to help us explore its regulatory mechanisms. In study, we reported comprehensive genome-wide DNA methylation landscape chicken cells, transcriptomic dynamics also presented. By uncovering patterns on individual genes, some genes accurately modulated by...

Traditional approaches for generating goat pluripotent stem cells (iPSCs) suffer from complexity and low preparation efficiency. Therefore, we tried to derive iPSCs with a new method by transfecting exogenous Oct4, Sox2, Klf4 c-Myc mRNAs into embryonic fibroblasts (GEFs), explore the mechanisms regarding transcription regulation of reprogramming factors in induction. four were transfected GEFs, localized nucleus approximately 90% transfection After five consecutive transfections, GEFs tended...

The use of stem cells in generating cell-based pacemaker therapies for bradyarrhythmia is currently being considered. Due to the propensity form tumors, as well ethical issues surrounding their use, seed used cardiac biological pacemakers have limitations. Very small embryonic-like (VSELs) are a unique and rare adult cell population, which same structural, genetic, biochemical, functional characteristics embryonic without controversy. In this study, we investigated ability rat bone marrow-...

ABSTRACT Stra8 (stimulated by retinoic acid gene 8) is a specific that expressed in mammalian germ cells during transition from mitosis to meiosis and plays key role the initiation of mammals birds. So, evaluation pathway cSSCs may provide deeper insight into spermatogenesis. miRNA was also an important regulating factor for SSCs. However, there currently no data indicating regulate SSCs via Stra8. Here, we predicted prospective targeting using online Bioinformatics database‐Targetscan,...

This study demonstrated different effects of bone morphogenetic protein 4 (BMP4) and retinoic acid (RA) signaling on the induction germ cell formation in chickens.

The CREPT (cell cycle-related and expression elevated protein in tumor, also known as RPRD1B) p15RS (p15INK4b -related sequence, RPRD1A) have been shown to regulate cell proliferation alter the cycle through Wnt/β-catenin pathway downstream genes human. Although several studies revealed mechanism by which human mammals, it is still unclear how these function poultry. In order determine of chicken, we examined a variety chicken tissues DF-1 cells. Then, determined effect overexpression or...

The present study investigated the dynamic expression and regulatory mechanism of transforming growth factor β (TGF-β) signaling involved in embryonic stem cells (ESCs) differentiation into male germ cells. Candidate genes TGF-β pathway were screened from RNA-sequencing (RNA-seq), which further validated by quantitative real-time PCR (qRT-PCR). Bone morphogenetic protein 4 (BMP4) was used to induce ESCs vitro. Inhibition reflected Western blot SMAD2 SMAD5 expression. Differentiating...

Germ cells have an irreplaceable role in transmitting genetic information from one generation to the next, and also play important sex differentiation poultry, while little is known about epigenetic factors that regulate germ cell differentiation. In this study, RNA-seq was used detect expression profiles of long non-coding RNAs (lncRNAs) during chicken embryonic stem (ESCs) into spermatogonial (SSCs). The results showed a total 296, 280 357 differentially expressed lncRNAs (DELs) were...

Abstract Currently, studies have analyzed the formation mechanism of primordial germ cell (PGC) at transcriptional level, but few protein which made study PGC not systematic. Here, we screened differential expression proteins (DEPs) regulated by label‐free proteomics with a novel sampling strategy embryonic stem cells and PGC. Analysis DEPs showed that multiple key events were involved, such as transition from glycolysis to oxidative phosphorylation, activation autophagy, low DNA methylation...

Autophagy plays an important role in the pluripotency and differentiation of stem cells. Transcriptome data showed that autophagy genes MAP1LC3A MAP1LC3B were significantly upregulated primordial germ cells (PGCs). The Kyoto Encyclopedia Genes Genome (KEGG) results lysosome signaling pathway, which is related to autophagy, was enriched PGCs. Quantitative RT-PCR, western blotting, transmission electron microscopy (TEM) expressed both embryonic (ESCs) PGCs but activated To explore chicken ESCs...

Embryonic stem cells (ESCs) are remarkably undifferentiated that originate from the inner cell mass of blastocyst. They possess ability to self-renew and differentiate into multiple types, making them invaluable in diverse applications such as disease modeling creation transgenic animals. In recent years, agricultural practices have evolved traditional biological breeding, it has become clear pluripotent (PSCs), either ESCs or induced (iPSCs), optimal for continually screening suitable...

The determination of sex in mammals is established and controlled by various complex mechanisms. In contrast, control poultry remains an unresolved issue. this study, RNA-sequencing was conducted for male gonads ovarian tissues chicken embryos up to 18.5 days identify metabolic factors influencing female differentiation, as well gonadal development. Our results reveal that

Chicken (Gallus gallus), as the most economically important poultry, is a classical and ideal model for studying mechanism of vertebrate developmental biology embryology. However, sex determination differentiation in chicken still elusive, which limited application slowed down many basic studies chicken. We applied PacBio Iso-seq to multiple spatiotemporal embryo-gonad tissues male female chicken, contain blastoderm (E0, un-differentiation stage), genital ridge (E3.5–6.5, sex-differentiation...

Abstract The study aims to analyze the key signaling pathways in regulating process of embryonic stem cells (ESCs) differentiation into spermatogonial (SSCs). We explored specific mechanisms C‐Jun amino‐terminal kinase (JNK) this process. Interference/overexpression MAPK8 allows JNK pathway be blocked/activated. In Retinoic acid (RA) induced vitro assays, expression germ cell marker genes, cvh, c‐kit, integrin α6 and β1 , was observed upregulate while activating significantly. Fluorescence...

Although many marker genes for germ cell differentiation have been identified, that specifically regulate primordial (PGC) generation are more difficult to determine. In the current study, we confirmed C2EIP is a PGC gene regulates by influencing expression of pluripotency-associated such as Oct4 and Sox2. Knockout during embryonic development reduced efficiency 1.5-fold, whereas overexpression nearly doubled both in vitro vivo. encodes cytoplasmic protein interacted with PTCH2 at...