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Simple Summary: The BMPR1B and T/Brachyury genes are considered major genetic factors influencing sheep reproduction and tail bone number. In this study, we conducted an association analysis between nine mutations of the BMPR1B gene and the litter sizes of short-tailed Gobi sheep and larger-tailed Ujimqin sheep. We also compared the frequency of the favorable alleles of four mutations among Gobi short tail sheep, Ujimqin sheep, and Mongolia sheep. In addition, we confirmed that the frequency of tail-bone-number-related T alleles at position c.363G>T in the T gene was significantly higher in short-tailed Gobi sheep than larger-tailed Ujimqin sheep. The significant deposition of tail fat in sheep has a profound impact on the economic benefits of animal husbandry. Furthermore, increasing the litter size is a crucial means of enhancing economic benefits. The BMPR1B and T/Brachyury genes are considered major functional genes that could affect sheep litter size and tail bone number, respectively. In this study, we employed direct sequencing to identify specific mutations of the BMPR1B gene in Gobi short tail sheep and carried out genotyping using MassARRAY technology for each variant of both the BMPR1B and T genes. Significant associations were demonstrated between the c.687G>A mutation of BMPR1B and the litter size in both the Gobi short tail sheep and Ujimqin sheep breeds. Meanwhile, the g.30058882_30058873GCAGATTAAAIndel mutation was significantly associated with the litter size in Gobi short tail sheep. These findings may provide valuable genetic markers for expanding sheep litter size. In addition, we also confirmed that the frequency of tail-bone-number-related T alleles was significantly higher in Gobi short tail sheep than in longer-tailed Ujimqin sheep. [ABSTRACT FROM AUTHOR]

Recent advances in understanding the CRISPR/Cas9 system have provided a precise and versatile approach for genome editing in various species. However, no study has reported simultaneous knockout of endogenous genes and site‐specific knockin of exogenous genes in large animal models. Using the CRISPR/Cas9 system, this study specifically inserted the fat‐1 gene into the goat MSTN locus, thereby achieving simultaneous fat‐1 insertion and MSTN mutation. We introduced the Cas9, MSTN knockout small guide RNA and fat‐1 knockin vectors into goat fetal fibroblasts by electroporation, and obtained a total of 156 positive clonal cell lines. PCR and sequencing were performed for identification. Of the 156 clonal strains, 40 (25.6%) had simultaneous MSTN knockout and fat‐1 insertion at the MSTN locus without drug selection, and 55 (35.25%) and 101 (67.3%) had MSTN mutations and fat‐1 insertions, respectively. We generated a site‐specific knockin Arbas cashmere goat model using a combination of CRISPR/Cas9 and somatic cell nuclear transfer for the first time. For biosafety, we mainly focused on unmarked and non‐resistant gene screening, and point‐specific gene editing. The results showed that simultaneous editing of the two genes (simultaneous knockout and knockin) was achieved in large animals, demonstrating that the CRISPR/Cas9 system has the potential to become an important and applicable gene engineering tool in safe animal breeding. [ABSTRACT FROM AUTHOR]

In this paper, catalytic wet oxidation is adopted to treat oilfield wastewater. Central composite design (CCD), an experimental design for response surface methodology (RSM), was used to create a set of 30 experimental runs needed for optimizing of the operating conditions. Temperature, catalyst quality, reaction time and oxidant dosage are examined factors. The experimental results show that catalytic wet oxidation could effectively reduce COD removal by 98.37%. Catalytic wet oxidation process is possibly suitable for a primary treatment for oilfield wastewater. [ABSTRACT FROM AUTHOR]

Simple Summary: Cashmere goats have double coats consisting of non-modulated fine inner hairs or cashmere fibers produced by secondary hair follicles (SHFs) and guard hairs produced by primary hair follicles (PHFs). Cashmere is an important economic product worldwide. The world market for cashmere is increasing while the current production of cashmere is limited. Thymosin beta-4 (Tβ4), a 4.9 kDa protein, contains 43 amino acids. Here, we investigated whether Tβ4 overexpression would increase SHF numbers, and thus improve the cashmere yield. We produced Tβ4 transgenic goats using a tissue-specific overexpression strategy. The Tβ4 overexpression (Tβ4-OE) goats had increased hair follicle development and higher cashmere yields than the wild type (WT) goats. The development of this goat model is not only valuable as a framework for future studies of the mechanism of goat follicular development, but will also lead to the improvement of economically important cashmere traits. Increased cashmere yield and improved quality are some goals of cashmere goat breeding. Thymosin beta-4 (Tβ4) plays a key role in the growth and development of hair follicles. For the past ten years, we have evaluated the role of Tβ4 by establishing a flock of 15 cashmere goats that specifically overexpress the Tβ4 gene in the hair follicles. These Tβ4 overexpression (Tβ4-OE) cashmere goats had more secondary hair follicles than the WT goats and produced more cashmere. Meanwhile, combined analysis of the skin transcriptome and proteome in cashmere goats suggested that Tβ4 may affect hair growth by interacting with keratin type II cytoskeletal 4 epidermal (KRT4) to mediate the extracellular signal-regulated protein kinase (ERK) signaling pathway, thereby promoting the development of secondary hair follicles, and consequently, increasing cashmere yield. Thus, the specific overexpression of Tβ4 in the hair follicles of cashmere goats effectively increased the cashmere yield. [ABSTRACT FROM AUTHOR]

Fibroblast growth factors (FGFs) are multifunctional signal molecules between cells, regulating the various physiological functions of the organism. FGF21 is a regulatory factor of the FGF family and has been postulated to play important roles in hair follicle development and hair follicle growth cycle. To evaluate the roles of FGF21, we had established a FGF21 knockout mouse model, using the CRISPR/Cas9 technology. We had constructed a FGF21 targeting vector and microinjected it with Cas9 mRNA and gRNA into fertilized ova of FVB mice. The gRNA was designed to target the exon 1 of the endogenous mouse FGF21 gene. Three lines of Fgf21 -/- mice were obtained from these experiments, and confirmed to harbor Fgf21 -/- genotypes and null expression phenotype, using DNA sequencing, qRT-PCR and Western blotting. FGF21 mRNA and FGF21 protein were not detected in tissues of these Fgf21 -/- mice. Depilation and histochemistry analyses showed that the Fgf21 -/- mice had lower body weight, slower hair regrowth and poorer hair quantities and smaller hair follicles diameters, as compared to WT mice. The Fgf21 -/- mice reported here could provide a useful genetic model for future studies of FGF21 functions in hair follicle development and hair follicle growth cycle.

To obtain bovine adipose-derived stem cells (ADSCs), bovine ADSCs were digested in collagenase type I solution. The growth curve of ADSCs was checked by cell counting. Chromosome analysis was checked. The molecular markers of ADSCs were detected with immunofluorescence staining. The morphology of ADSCs was identical to fibroblast like and the cells showed active proliferative ability. Vimentin, CD49d and CD13 antigens were detected, but CD34 antigen was negative. Alkaline phosphatase activity was greater in ADSCs during calcification, and Alizarin Red staining was positive. Lipid droplets were apparent around cells during adipogenesis, and Oil Red-O staining was positive. The results demonstrated that ADSCs could be used as seed cells for tissue engineering due to the simple isolation, differentiation and stable and active growth.