Complexation of retroviruses with polymers significantly increases the number of genes transferred to murine embryonic stem cells, but does not raise transgene expression levels

Embryonic stem cells efficiently silence retrovirus transgene expression. To help to solve this problem, retroviruses have been developed that are more resistant to silencing, such as retroviruses derived from the MSCV (murine‐stem‐cell virus). A complementary approach to increasing transgene expres...

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Bibliographic Details
Published in:Biotechnology and applied biochemistry 2008-11, Vol.51 (3), p.141-151
Main Authors: Chilton, Jamie M., Le Doux, Joseph M.
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Biotechnology
Cattle
Cell Line
Cell Proliferation
Chondroitin Sulfates - metabolism
embryonic stem call
Embryonic Stem Cells - cytology
Embryonic Stem Cells - metabolism
Fundamental and applied biological sciences. Psychology
Gene Expression
Gene Transfer Techniques
Hexadimethrine Bromide - metabolism
Mice
murine-stem-cell virus (MSCV)
polymer
Polymers - metabolism
Retroviridae - metabolism
retrovirus
RNA, Messenger - genetics
RNA, Messenger - metabolism
Transduction, Genetic
transgene expression
Transgenes - genetics
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Summary:Embryonic stem cells efficiently silence retrovirus transgene expression. To help to solve this problem, retroviruses have been developed that are more resistant to silencing, such as retroviruses derived from the MSCV (murine‐stem‐cell virus). A complementary approach to increasing transgene expression might be to increase the number of integrated transgenes. To test this approach, we formed polymer complexes with MSCV‐derived ecotropic retroviruses, concentrated them up to 40‐fold and transduced two different murine embryonic stem cell lines, with a mouse fibroblast cell line as a control. The number of integrated transgenes increased more than 50‐fold in the embryonic stem cell lines, yet, surprisingly, transgene expression did not increase. Interestingly, the embryonic stem cells had significantly fewer integrated transgenes than the mouse fibroblasts, even though transduction conditions were identical, which suggests that embryonic stem cells may restrict a post‐binding step of retrovirus transduction.
ISSN:0885-4513
1470-8744
DOI:10.1042/BA20070213