Project description:Haploid androgenetic embryonic stem cells (haESCs) can produce live mice, thus have potential for genetic manipulation. However, practical application is limited by self-diploidization, loss-of-imprint, low-birth-rate, and livestock haESCs have yet to be achieved. Here we show that combination of FACE medium supports establishment of durable haESCs from cattle and sheep that overcome above problems. Multi-omics and functional analysis of FACE-haESCs showed that at a naive-to-primed intermediate pre-gastrulation formative state of pluripotency. Moreover, transient protamine overexpression remodels ruminant haESCs nuclei into spermatid-like structures concomitant with removal of ICHI barrier, resulting blastocyst rate of Pro-ICHI similar to that of in vitro fertilization (IVF) embryos. As a proof-of-concept, we also developed a novel Pro-ICHI::ePE system to fast produce trait-improved cattle with exogene-free effects.
Project description:Haploid androgenetic stem cells (haSCs) are invaluable resources for studying animal traits and greatly impact livestock breeding, but livestock haSCs have yet to be obtained. Here, we report the derivation of cattle and sheep haSCs using the recombined FACE medium, and these cells harbor formative pluripotency features. Notably, knockout cattle and sheep can be obtained upon injection of the protaminized haSCs with targeted gene mutations into intact oocytes.
Project description:Structural and functional impacts of copy number variations (CNVs) on livestock genomes are not yet well understood. In this study, we have identified 1853 CNV regions (CNVRs) using population-scale sequencing data generated from 75 cattle of 8 breeds (Holstein, Angus, Jersey, Limousin, Romagnola, Brahman, Gir and Nelore). Individual genome sequence coverage ranged from 4 to 30 fold, with a mean of 11.8 fold. A total of 3.1% (87.5 Mb) of the cattle genome is predicted to be copy number variable, representing a substantial increase over the previous estimates (~2%). This dataset was highly correlated with array CGH data (r2 = 0.761) and was validated to be accurate with an estimated 12% false positive rate and a 19% false negative rate based on qPCR and array CGH, respectively. Hundreds of CNVs were found to be either breed specific or differentially variable across breeds, including the RICTOR gene in dairy breeds and the PNPLA3 gene in the beef breeds. In contrast, clusters of the PRP and PAG genes are duplicated in all sequenced animals, implicating that subfunctionalization, neofunctionalization or overdominance play a role in diversifying these fertility related genes. Further population-genetic analyses based on CNVs revealed the population structures of these taurine and indicine breeds and uncovered hundreds of positively selected CNV candidates near important functional genes. These CNV results provide a new glimpse of diverse selections during cattle speciation, domestication, breed formation, and recent genetic improvement.
Project description:This project aimed to discover the protein-based biomarkers for tick resistance in cattle using cattle skin samples. The cattle were phenotyped into two groups, tick-resistant and susceptible after artificial tick challenge. Mean tick scores were used to categorise cattle. The SWATH analysis was sued to measure the relative abundance of proteins in skin samples of the two groups at different time points.
Project description:This project aimed to discover the protein-based biomarkers for tick resistance in cattle using cattle serum samples. The cattle were phenotyped into two groups, tick-resistant and susceptible after an artificial tick challenge. Mean tick scores were used to categorise cattle. The SWATH analysis was sued to measure the relative abundance of proteins in skin samples of the two groups at different time points.
Project description:This trial was undertaken to examine the perhipheral cellular and antibody response of cattle following infestation with the cattle tick, Rhipicephalus microplus. The information from the Affymetrix gene expression data is used to complement other measurements of immune function such as cellular subset composition and antibody response in cattle of high (Brahman) and low (Holstein-Friesian) resistance to the cattle tick. Keywords: Disease state analysis
Project description:Haploid androgenetic stem cells (haSCs) are invaluable resources for studying animal traits and greatly impact livestock breeding, but livestock haSCs have yet to be obtained. Here, we report the derivation of cattle and sheep haSCs using the recombined FACE medium, and these cells harbor formative pluripotency features. Notably, knockout cattle and sheep can be obtained upon injection of the protaminized haSCs with targeted gene mutations into intact oocytes.
Project description:Haploid androgenetic stem cells (haSCs) are invaluable resources for studying animal traits and greatly impact livestock breeding, but livestock haSCs have yet to be obtained. Here, we report the derivation of cattle and sheep haSCs using the recombined FACE medium, and these cells harbor formative pluripotency features. Notably, knockout cattle and sheep can be obtained upon injection of the protaminized haSCs with targeted gene mutations into intact oocytes.