scholarly journals 5’-flanking variants of equine casein genes (CSN1S1, CSN1S2, CSN2, CSN3) and their relationship with gene expression and milk composition

2018 ◽  
Vol 60 (1) ◽  
pp. 71-78 ◽  
Author(s):  
Jakub Cieslak ◽  
Lukasz Wodas ◽  
Alicja Borowska ◽  
Piotr Pawlak ◽  
Grazyna Czyzak-Runowska ◽  
...  
Keyword(s):  
Gene Expression ◽  
Milk Composition ◽  
Casein Genes

scholarly journals A hypomorphic mutation in the mouse Csn1s1 gene generated by CRISPR/Cas9 pronuclear microinjection

2021 ◽  
Vol 25 (3) ◽  
pp. 331-336
Author(s):  
A. V. Smirnov ◽  
T. A. Shnaider ◽  
A. N. Korablev ◽  
A. M. Yunusova ◽  
I. A. Serova ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Knockout ◽  
Milk Proteins ◽  
Transgenic Animals ◽  
Regulatory Elements ◽  
Milk Composition ◽  
Peptide Sequence ◽  
Start Codon ◽  
Signal Peptide Sequence ◽  
Casein Genes

Caseins are major milk proteins that have an evolutionarily conserved role in nutrition. Sequence variations in the casein genes affect milk composition in livestock species. Regulatory elements of the casein genes could be used to direct the expression of desired transgenes into the milk of transgenic animals. Dozens of casein alleles have been identified for goats, cows, sheep, camels and horses, and these sequence variants are associated with altered gene expression and milk protein content. Most of the known mutations affecting casein genes’ expression are located in the promoter and 3’-untranslated regions. We performed pronuclear microinjections with Cas9 mRNA and sgRNA against the first coding exon of the mouse Csn1s1 gene to introduce random mutations in the α-casein (Csn1s1) signal peptide sequence at the beginning of the mouse gene. Sanger sequencing of the founder mice identified 40 mutations. As expected, mutations clustered around the sgRNA cut site (3 bp from PAM). Most of the mutations represented small deletions (1–10 bp), but we detected several larger deletions as well (100–300 bp). Functionally most mutations led to gene knockout due to a frameshift or a start codon loss. Some of the mutations represented in-frame indels in the first coding exon. Of these, we describe a novel hypomorphic Csn1s1 (Csn1s1c.4-5insTCC) allele. We measured Csn1s1 protein levels and confirmed that the mutation has a negative effect on milk composition, which shows a 50 % reduction in gene expression and a 40–80 % decrease in Csn1s1 protein amount, compared to the wild-type allele. We assumed that mutation affected transcript stability or splicing by an unknown mechanism. This mutation can potentially serve as a genetic marker for low Csn1s1 expression.

Functional study of the equine β-casein and κ-casein gene promoters

2005 ◽  
Vol 72 (S1) ◽  
pp. 34-43 ◽  
Author(s):  
Tina Lenasi ◽  
Nadja Kokalj-Vokac ◽  
Mojca Narat ◽  
Antonella Baldi ◽  
Peter Dovc
Keyword(s):  
Gene Expression ◽  
Mammary Gland ◽  
Reporter Gene ◽  
Gene Promoter ◽  
Reporter Gene Expression ◽  
Proximal Promoter ◽  
Gene Promoters ◽  
Tissue Specific ◽  
Casein Gene ◽  
Casein Genes

Casein genes are expressed in a tissue-specific and highly coordinated manner. The main goals of casein gene promoter studies are to unravel cis- and trans-acting factors involved in the complex signalling pathway controlling milk production, and to explore the possibility of using these promoters for tissue-specific production of heterologous proteins in the mammary gland. Here we present a comparative study of the equine β-casein and κ-casein gene proximal promoters. In order to confirm the assumption that in the horse, as in other mammalian species, casein genes are organized in a cluster located on a single chromosome, we performed in situ hybridization of pro-metaphase chromosomes with two BAC clones containing different equine casein genes. Sequence analysis of the β-casein and κ-casein gene proximal promoters revealed binding sites for activators (STAT5, GRE, NF1, MAF) and repressors (YY1, PMF), characteristic for casein genes. The alignments of casein gene promoters revealed the highest sequence identity in the proximal promoter region between the equine and human β-casein gene promoters. We directly compared the activity of equine β-casein and κ-casein gene promoters in vitro using bovine mammary gland cell line BME-UV1. In this system, the κ-casein gene proximal promoter activated the reporter gene expression more efficiently than the β-casein gene promoter of approximately the same length. The 810 bp of β-casein promoter activated the reporter gene expression more efficiently than the long fragment (1920 bp) and the 1206 bp fragment of the same promoter, which included also 396 bp of 5′ UTR.

Gene transfer in animal production

1988 ◽  
Vol 12 ◽  
pp. 1-14
Author(s):  
A.J. Clark
Keyword(s):  
Gene Expression ◽  
Gene Transfer ◽  
Molecular Level ◽  
Milk Composition ◽  
Farm Animals ◽  
Research Groups ◽  
Genetic Changes ◽  
Pharmaceutical Proteins ◽  
Pronuclear Injection ◽  
Recent Developments

ABSTRACTGene transfer by pronuclear injection has been accomplished in farm animals by a number of research groups. Applications of this technology for improving milk composition, producing pharmaceutical proteins and manipulating physiology are described. Recent developments in our understanding of gene expression at the molecular level will increase the precision with which genetic changes can be made by gene transfer.

scholarly journals Activation of Adipocyte mTORC1 Increases Milk Lipids in a Mouse Model of Lactation

2021 ◽  
Author(s):  
Noura El-Habbal ◽  
Allison C. Meyer ◽  
Hannah Hafner ◽  
JeAnna R. Redd ◽  
Zach Carlson ◽  
...  
Keyword(s):  
Gene Expression ◽  
Fatty Acids ◽  
Mammary Gland ◽  
Milk Fat ◽  
Saturated Fatty Acids ◽  
Milk Composition ◽  
Nutrient Sensing ◽  
Lower Percentage ◽  
Milk Lipids

Human milk is the recommended nutrient source for newborns. The mammary gland comprises multiple cell types including epithelial cells and adipocytes. The contributions of mammary adipocytes to breast milk composition and the intersections between mammary nutrient sensing and milk lipids are not fully understood. A major nutrient sensor in most tissues is the mechanistic target of rapamycin 1 (mTORC1). To assess the role of excess nutrient sensing on mammary gland structure, function, milk composition, and offspring weights, we used an Adiponectin-Cre driven Tsc1 knockout model of adipocyte mTORC1 hyperactivation. Our results show that the knockout dams have higher milk fat contributing to higher milk caloric density and heavier offspring weight during lactation. Additionally, milk of knockout dams displayed a lower percentage of saturated fatty acids, higher percentage of monounsaturated fatty acids, and a lower milk ω6: ω3 ratio driven by increases in Docosahexaenoic acid (DHA). Mammary gland gene expression analyses identified changes in eicosanoid metabolism, adaptive immune function and contractile gene expression. Together, these results suggest a novel role of adipocyte mTORC1 in mammary gland function and morphology, milk composition, and offspring growth.

Evaluation of xanthosine treatment on gene expression of mammary glands in early lactating goats

2018 ◽  
Vol 85 (3) ◽  
pp. 288-294 ◽  
Author(s):  
Ratan K Choudhary ◽  
Shanti Choudhary ◽  
Devendra Pathak ◽  
Rahul Udehiya ◽  
Ramneek Verma ◽  
...  
Keyword(s):  
Gene Expression ◽  
Milk Yield ◽  
Progenitor Cell ◽  
Milk Composition ◽  
Specific Gene ◽  
Dairy Goats ◽  
Differentiation Markers ◽  
Cell Markers ◽  
Lactating Goats ◽  
Mammary Stem

This study examined the hypothesis that xanthosine (XS) treatment would promote mammary-specific gene expression and stem cell transcripts and have a positive influence on milk yield of dairy goats. Seven primiparous Beetal goats were assigned to the study. Five days after kidding, one gland (either left or right) was infused with XS (TRT) twice daily for 3 d and the other gland with no XS infusion served as a control (CON). Mammary biopsies were collected at 10 d and RNA was isolated. Gene expression analysis of milk synthesis genes, mammary stem/progenitor cell markers, cell proliferation and differentiation markers were performed using real time quantitative PCR (RT-qPCR). Results showed that the transcripts of milk synthesis genes (BLG4, CSN2, LALBA, FABP3, CD36) and mammary stem/progenitor cell markers (ALDH1 and NR5A2) were increased in as a result of XS treatment. Average milk yield in TRT glands was increased marginally (approximately ~2% P = 0·05, paired t-test) per gland relative to CON gland until 7 wk. After 7 wk, milk yield of TRT and CON glands did not differ. Analysis of milk composition revealed that protein, lactose, fat and solids-not-fat percentages remained the same in TRT and CON glands. These results suggest that XS increases expression of milk synthesis genes, mammary stem/progenitor cells and has a small effect on milk yield.

scholarly journals Effect of β-lactoglobulin and κ-casein genes polymorphism on milk composition in indigenous Zel sheep

2013 ◽  
Vol 56 (1) ◽  
pp. 216-224 ◽  
Author(s):  
S. Yousefi ◽  
M. A. Azari ◽  
S. Zerehdaran ◽  
R. Samiee ◽  
R. Khataminejhad
Keyword(s):  
Gene Polymorphism ◽  
Pcr Amplification ◽  
Milk Composition ◽  
Sheep Breed ◽  
Pcr Products ◽  
Casein Genes ◽  
Β Lactoglobulin

Abstract. The effect of β-lactoglobulin (β-LG) and κ-casein (CSN3) genotypes on milk composition were evaluated in Iranian indigenous Zel sheep breed. Genotypes were determined by PCR amplification followed by digestion with RsaI enzyme for exon II of the β-LG gene and SSCP method for exon IV of the CSN3 gene. Polymorphism was detected in all PCR products. β-lactoglobulin showed two alleles and three genotypes and CSN3 gene revealed two conformational patterns, respectively. Results indicated that there were significant associations between AB genotype of β-LG gene with higher fat and lactose percentages and also between K1 pattern of CSN3 gene with higher lactose percentage. Therefore, it is feasible to improve milk composition in Zel sheep breed using β-LG and CSN3 genes.

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