In:
Stem Cells International, Hindawi Limited, Vol. 2011 ( 2011), p. 1-14
Abstract:
Non-viral integrating systems, PhiC31 phage integrase ( ϕ C31), and Sleeping Beauty transposase (SB), provide an effective method for ex vivo gene delivery into cells. Here, we used a plasmid-encoding GFP and neomycin phosphotransferase along with recognition sequences for both ϕ C31 and SB integrating systems to demonstrate that both systems effectively mediated integration in cultured human fibroblasts and in rat multipotent adult progenitor cells (rMAPC). Southern blot analysis of G418-resistant rMAPC clones showed a 2-fold higher number of SB-mediated insertions per clone compared to ϕ C31. Sequence identification of chromosomal junction sites indicated a random profile for SB-mediated integrants and a more restricted profile for ϕ C31 integrants. Transgenic rMAPC generated with both systems maintained their ability to differentiate into liver and endothelium albeit with marked attenuation of GFP expression. We conclude that both SB and ϕ C31 are effective non-viral integrating systems for genetic engineering of MAPC in basic studies of stem cell biology.
Type of Medium:
Online Resource
ISSN:
1687-966X
,
1687-9678
Language:
English
Publisher:
Hindawi Limited
Publication Date:
2011
detail.hit.zdb_id:
2573856-2