Somatic cell clones often fail at a developmental stage coincident with commencement of differentiation. The transcription factor Oct4 is expressed during cleavage stages and essential for the differentiation blastocyst. expression becomes restricted to inner mass epiblast. After gastrulation active only in germ cells silent somatic cells. Here, an Oct4-GFP transgene were used as markers which gene reprogramming could be directly related potential clones. Cumulus initiated correct but showed...

Piwi-interacting RNAs (piRNAs) are essential for silencing of transposable elements in the germline, but their biogenesis is poorly understood. Here we demonstrate that MOV10L1, a germ cell–specific putative RNA helicase, associated with Piwi proteins. Genetic disruption MOV10L1 helicase domain mice renders both MILI and MIWI2 devoid piRNAs. Absence functional piRNA pathway Mov10l1 mutant testes causes loss DNA methylation subsequent derepression retrotransposons cells. The males sterile...

Meiotic silencing of sex chromosomes may cause their depletion meiosis-specific genes during evolution. Here, we challenge this hypothesis by reporting the identification TEX11 as first X-encoded factor in mice. forms discrete foci on synapsed regions meiotic and appears to be a novel constituent nodules involved recombination. Loss function causes chromosomal asynapsis reduced crossover formation, leading elimination spermatocytes, respectively, at pachytene anaphase I stages. Specifically,...

Gene editing of the mitochondrial genome using CRISPR-Cas9 system is highly challenging mainly due to sub-efficient delivery guide RNA and Cas9 enzyme complexes into mitochondria. In this study, we were able perform gene in DNA by appending an NADH-ubiquinone oxidoreductase chain 4 (ND4) targeting transport-derived stem loop element (RP-loop) expressing with a preceding localization sequence. We observe colocalization RP-loop gRNA marked reduction ND4 expression cells carrying 11205G variant...

During meiosis, homologous chromosomes undergo synapsis and recombination. We identify TEX15 as a novel protein that is required for chromosomal meiotic Loss of function in mice causes early arrest males but not females. Specifically, TEX15-deficient spermatocytes exhibit failure synapsis. In mutant spermatocytes, DNA double-strand breaks (DSBs) are formed, localization the recombination proteins RAD51 DMC1 to severely impaired. Based on these data, we propose regulates loading repair onto...

The POU-domain transcription factor Pou5f1 (Oct4) is restricted to pluripotent embryonic cells and the germ line of mouse required for maintenance pluripotency within inner cell mass blastocyst. Despite highly conserved genomic organization regulatory regions between Oct4 gene its bovine orthologue, protein not blastocyst-stage embryos, suggesting that may be a key regulator in bovine. We analyze temporal spatial distribution transcript oocytes preimplantation-stage contrast distribution, we...

A micro/nano-fabrication process of a nanochannel electroporation (NEP) array and its application for precise delivery plasmid non-viral gene transfection is described. dip-combing device optimized to produce DNA nanowires across microridge patterned on the polydimethylsiloxane (PDMS) surface with yield up 95%. Molecular imprinting based low viscosity resin, 1,4-butanediol diacrylate (1,4-BDDA), adopted convert microridge-nanowire-microridge into microchannel-nanochannel-microchannel (MNM)...

Besides holding great promise in clinics, embryonic stem (ES) cells represent a valuable tool for studying regulation of early developmental processes, such as cell differentiation preimplantation embryos. The caudal-related homeobox protein Cdx2 is transcriptional regulator essential trophoblast lineage, functioning implantation. Using an inducible system, we show that gain function ES triggers trophoblast-like morphological differentiation, accompanied by ploidy increase, onset expression...

Parthenogenetic embryonic stem (ES) cells with two oocyte-derived genomes (uniparental) have been proposed as a source of autologous tissue for transplantation. The therapeutic applicability any uniparental cell type is uncertain due to the consequences genomic imprinting that in mammalian tissues causes unbalanced expression imprinted genes. We transplanted fetal liver into lethally irradiated adult mice test their capacity replace hematopoietic tissue. Both maternal (gynogenetic) and...

Histone H1 is an abundant and essential component of chromatin whose precise role in regulating gene expression poorly understood.Here, we report that a major target H1-mediated regulation embryonic stem (ES) cells the X-linked Rhox homeobox cluster.To address underlying mechanism, examined founding member cluster-Rhox5-and found its distal promoter (Pd) loses H1, undergoes demethylation, transcriptionally activated response to loss genes ES cells.Demethylation Pd required for...

Ubiquitin E3 ligases target their substrates for ubiquitination, leading to proteasome-mediated degradation or altered biochemical properties. The ubiquitin ligase Ubr2, a recognition component of the N-end rule proteolytic pathway, recognizes proteins with N-terminal destabilizing residues and plays an important role in spermatogenesis. Tex19.1 (also known as Tex19) has been previously identified germ cell-specific protein mouse testis. Here we report that forms stable complex Ubr2 testes....

Parthenogenesis is the development of an oocyte without fertilization. Mammalian parthenogenetic (PG) embryos are not viable, but can develop into blastocysts from which embryonic stem cells (ESCs) have been derived in mouse and human. PG ESCs frequently homozygous for alleles encoding major histocompatibility complex (MHC) molecules. MHC homozygosity permits much more efficient immune matching than heterozygosity found conventional ESCs, making a promising cell source therapies requiring no...

Tudor domain containing (Tdrd) proteins that are expressed in germ cells divided into two groups. One group, consisting of TDRD1, TDRKH, TDRD9 and TDRD12, function piRNA biogenesis retrotransposon silencing, while the other group including RNF17/TDRD4 TDRD5-7 required for spermiogenesis. These Tdrd play distinct roles during male cell development. Here, we report characterization STK31/TDRD8 mice. STK31 contains a tudor serine/threonine kinase domain. We find is cytoplasmic protein cells....

Parent-of-origin imprints have been implicated in the regulation of neural differentiation and brain development. Previously we shown that, despite lack a paternal genome, human parthenogenetic (PG) embryonic stem cells (hESCs) can form proliferating (NSCs) that are capable into physiologically functional neurons while maintaining allele-specific expression imprinted genes. Since biparental (“normal”) hESC-derived NSCs (N NSCs) targeted by immune cells, characterized immunogenicity PG NSCs....

Uniparental zygotes with two paternal (androgenetic [AG]) or maternal (gynogenetic [GG]; parthenogenetic [PG]) genomes are not able to develop into viable offspring but can form blastocysts from which embryonic stem cells (ESCs) be derived. Although some aspects of the in vitro and vivo differentiation potential PG GG ESCs several species have been studied, developmental capacity AG is much less clear. Here, we investigate murine undergo neural differentiation. We observed that differentiate...

Parent of origin imprints on the genome have been implicated in regulation neural cell type differentiation. The ability human parthenogenetic (PG) embryonic stem cells (hpESCs) to undergo lineage and type-specific differentiation is undefined. We determined potential hpESCs differentiate into various subtypes. Concurrently, we examined DNA methylation expression status imprinted genes. Under culture conditions promoting differentiation, hpESC-derived (hpNSCs) gave rise glia neuron-like that...

In eukaryotes, mRNA is actively exported to the cytoplasm by a family of nuclear RNA export factors (NXF). Four Nxf genes have been identified in mouse: Nxf1, Nxf2, Nxf3, and Nxf7. Inactivation germ cell-specific gene, causes defects spermatogenesis. Here we report that Nxf3 expressed exclusively Sertoli cells postnatal testis, developmentally regulated manner. Expression coincides with cessation cell proliferation beginning their differentiation. Continued expression mature adult...

ABSTRACT Gene editing of the mitochondrial genome using CRISPR-Cas9 system is highly challenging mainly due to sub-efficient delivery guide RNA and Cas9 enzyme complexes into mitochondria. In this study, we were able perform gene in DNA by appending NADH-ubiquinone oxidoreductase chain 4 (ND4) targeting a transport derived stem loop element (RP-loop) expressing with preceding localization sequence. Our results showed co-localization RP-loop gRNA marked reduction ND4 expression cells carrying...

The inefficiency of mammalian somatic cell cloning is associated with abnormal gene expression presumably caused by errors in reprogramming the transplanted genome. In mouse, aggregation four-cell stage clones leads to an improvement both and development. To determine whether clone-clone at postgenomic activation stages influences bovine clones, we profiled, single aggregated embryos blastocyst stage, developmentally relevant genes namely Oct4, Dnmt1, Dnmt3, Glut1, Glut3, a housekeeping...

Mammalian somatic cell cloning requires factors specific to the oocyte for reprogramming succeed. This does not exclude that continues during zygote and cleavage stages. The capacity or role of zygotic stages reprogram nuclei is difficult assess due limited development transplanted into cytoplasts these Alternatively, tetraploid embryos have been used study can be assessed their contribution extra-embryonic lineages. When mouse cumulus transgenic Oct4-green fluorescent protein (GFP) were...