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A severe de novo methylation of episomal vectors by human ES cells.

Abstract：

:Episomal vectors can allow efficient genetic modification of cells and have the potential advantage of avoiding chromosomal position of integration effects. Here we explore the use of an Epstein-Barr virus-based episomal vector with human embryonic stem (ES) cells, and find high initial transfection rates, but a rapid loss of reporter gene expression. Similar to mouse ES cells, human ES cells express high levels of the de novo DNA methyltransferases, and we detected dramatic CpG methylation and minor non-CpG methylation on the episomes recovered from the human ES cells 7 days after the transfection, which was not present on the same episome recovered from 293 cells. Interestingly, the oriP region of the episomes was relatively excluded from this methylation. These findings define some of the limitations of using episomal vectors with human ES cells and offer a unique platform for analyzing epigenetic gene silencing in human ES cells.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Kameda T,Smuga-Otto K,Thomson JA

doi

10.1016/j.bbrc.2006.08.175

subject

Has Abstract

pub_date

2006-11-03 00:00:00

pages

1269-77

issue

4

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(06)01987-5

journal_volume

349

pub_type

杂志文章

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