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Bacterial artificial chromosomes establish replication timing and sub-nuclear compartment de novo as extra-chromosomal vectors [4C]

ABSTRACT: The role of DNA sequence in determining replication timing (RT) and chromatin higher order organization remains elusive. To address this question, we have developed an extra-chromosomal replication system consisting of ~200kb human bacteria artificial chromosomes (BACs) modified with Epstein-Barr virus (EBV) replication origin elements (E-BACs). E-BACs were stably maintained as autonomous mini-chromosomes in both HeLa and human induced pluripotent stem cells (hiPSCs) and established their RT de novo. We applied 4C-seq to evaluate E-BACs' sub-nuclear compartment.

ORGANISM(S): Homo sapiens

PROVIDER: GSE102521 | GEO | 2018/01/29

REPOSITORIES: GEO

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Bacterial artificial chromosomes establish replication timing and sub-nuclear compartment de novo as extra-chromosomal vectors [repli-seq]

Project description:The role of DNA sequence in determining replication timing (RT) and chromatin higher order organization remains elusive. To address this question, we have developed an extra-chromosomal replication system consisting of ~200kb human bacteria artificial chromosomes (BACs) modified with Epstein-Barr virus (EBV) replication origin elements (E-BACs). E-BACs were stably maintained as autonomous mini-chromosomes in both HeLa and human induced pluripotent stem cells (hiPSCs) and established their RT de novo. We applied repli-seq to evaluate E-BACs' replication timing.

2018-01-29 | GSE102522 | GEO

Bacterial artificial chromosomes establish replication timing and sub-nuclear compartment de novo as extra-chromosomal vectors.

Project description: Not available

| S-EPMC5829748 | biostudies-literature

Bacterial artificial chromosomes establish replication timing and sub-nuclear compartment de novo as extra-chromosomal vectors

Project description:Bacterial artificial chromosomes establish replication timing and sub-nuclear compartment de novo as extra-chromosomal vectors

| PRJNA429696 | ENA

Bacterial artificial chromosomes establish replication timing and sub-nuclear compartment de novo as extra-chromosomal vectors

Project description:Bacterial artificial chromosomes establish replication timing and sub-nuclear compartment de novo as extra-chromosomal vectors

| PRJNA398470 | ENA

Bacterial artificial chromosomes establish replication timing and sub-nuclear compartment de novo as extra-chromosomal vectors

Project description:This SuperSeries is composed of the SubSeries listed below.

2018-01-29 | GSE102704 | GEO

Uniform, integration-free iPS cells with a safeguard system using human artificial chromosome vectors

Project description:Human artificial chromosomes (HACs) have unique characteristics as gene-delivery vector, e.g., episomal transmission and transfer of multiple, large transgenes. Here, we demonstrate the advantages of HAC vectors for reprogramming mouse embryonic fibroblasts (MEFs) to induced pluripotent stem (iPS) cells. Two HAC vectors (iHAC1 and iHAC2) were constructed. Both carried four reprogramming factors, and iHAC2 also encoded a p53-knockdown construct (see Kazuki et al 2011). The iHAC1 partially reprogrammed MEFs, and the iHAC2 efficiently reprogrammed MEFs. Global gene expression showed that the iHACs, unlike other vectors, generated relatively uniform iPS cells. We established iHAC-free iPS cells by isolating cells that spontaneously lost the iHAC2. Analyses of pluripotent markers, teratomas, and chimeras confirmed that these iHAC-free iPS cells were pluripotent. Moreover, iHAC-free iPS cells with a re-introduced HAC encoding Herpes Simplex Virus Thymidine Kinase were eliminated by Ganciclovir treatment; therefore, the HAC safeguard system functioned in iPS cells. Thus, the HAC vector could generate uniform, integration-free iPS cells with a safeguard system.

2011-10-16 | GSE29441 | GEO

Uniform, integration-free iPS cells with a safeguard system using human artificial chromosome vectors

2011-10-15 | E-GEOD-29441 | biostudies-arrayexpress

Regions of XY homology in the pig X chromosome and the boundary of the pseudoautosomal region

Project description:Background Sex chromosomes are subject to evolutionary pressures distinct from the remainder of the genome, shaping their structure and sequence content. We are interested in the sex chromosomes of domestic pigs (Sus scrofa), how their structure and gene content compares and contrasts with other mammalian species, and the role of gonosomal genes in fertility. This requires an understanding of the XY-homologous sequence on these chromosomes. To this end, we performed microarray-based comparative genomic hybridisation (array-CGH) with male and female Duroc genomic DNA on a pig X-chromosome BAC tiling-path microarray. Putative XY-homologous BACs from regions of interest were subsequently FISH mapped. Results We show that the porcine PAR is approximately 6.5-6.9Mb at the beginning of the short arm of the X, with gene content reflective of the artiodactyl common ancestor. Our array-CGH data also shows an XY-homologous region close to the end of the X long arm, spanning three X BACs. These BACs were FISH mapped, and paint the entire long arm of SSCY. Further clones of interest revealed X-autosomal homology or regions containing repetitive content. Conclusions This study has identified regions of XY homology in the pig genome, and defined the boundary of the PAR on the X chromosome. This adds to our understanding of the evolution of the sex chromosomes in different mammalian lineages, and will prove valuable for future comparative genomic work in suids and for the construction and annotation of the genome sequence for the sex chromosomes. Our finding that the SSCYq repetitive content has corresponding sequence on the X chromosome gives further insight into structure of SSCY, and suggests further functionally important sequences remain to be discovered on the X and Y.

2012-08-30 | GSE40244 | GEO

Exogenous artificial DNA forms chromatin structure with active transcription in yeast [ChIP-seq]

Project description:Chromosomes and plasmids are two forms of genetic carriers. Exogenous yeast artificial chromosomes are also considered as yeast centromeric plasmids in many cases. Here, we used state-of-the-art sequencing technologies to comprehensively profile the genetic, epigenetic, transcriptional and proteomic characteristics of an exogenous data-carrying chromosome. We found that the data-carrying DNA formed active chromatin with high chromatin accessibility and H3K4 tri-methylation levels. We also confirmed that the data-carrying chromosome had a circular shape in the nucleus and was arranged in the Rabl configuration, which may contribute to the self-replication and haploidy of the chromosome in vivo. The data-carrying chromosome displayed highly pervasive transcriptional ability and transcribed hundreds of non-coding RNAs. In summary, this work explores the chromatin epigenetic state, chromatin structure and transcriptional landscape of an exogenous artificial chromosome. The results demonstrated that the exogenous artificial chromosome did form a chromatin structure and was not a naked and incompact plasmid, which strengthen our understanding of artificial chromosomes.

2022-09-30 | GSE183491 | GEO

Exogenous artificial DNA forms chromatin structure with active transcription in yeast [RNA-seq]

2022-09-30 | GSE183490 | GEO

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