Episomal minicircles persist in periods of transcriptional inactivity and can be transmitted through somatic cell nuclear transfer into bovine embryos View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2019-01-29

AUTHORS

Stefan Wagner, Judi McCracken, Sabine Bruszies, Ric Broadhurst, David N. Wells, Björn Oback, Jürgen Bode, Götz Laible

ABSTRACT

Episomal plasmids based on a scaffold/matrix attachment region (S/MAR) are extrachromosomal DNA entities that replicate once per cell cycle and are stably maintained in cells or tissue. We generated minicircles, episomal plasmids devoid of bacterial sequences, and show that they are stably transmitted in clonal primary bovine fibroblasts without selection pressure over more than two months. Total DNA, plasmid extraction and fluorescence in situ hybridization (FISH) analyses suggest that the minicircles remained episomal and were not integrated into the genome. Minicircles survived extended periods in serum-starved cells, which indicates that ongoing transcription in non-proliferating cells is not necessary for the maintenance of S/MAR-episomes. To test whether minicircles endure the process of somatic cell nuclear transfer (SCNT), we used cell-cycle synchronized, serum-starved, minicircle-containing cells. Analysis of cells outgrown from SCNT-derived blastocysts shows that the minicircles are maintained through SCNT and early embryonic development, which raises the prospect of using cell lines with episomal minicircles for the generation of transgenic animals. More... »

PAGES

1-10

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s11033-019-04624-x

DOI

http://dx.doi.org/10.1007/s11033-019-04624-x

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1111764491

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/30694456


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