PhiC31 integrase induces a DNA damage response and chromosomal rearrangements in human adult fibroblasts
(2009) In BMC Biotechnology 9.- Abstract
- Background: PhiC31 integrase facilitates efficient integration of transgenes into human and mouse genomes and is considered for clinical gene therapy. However recent studies have shown that the enzyme can induce various chromosomal abnormalities in primary human embryonic cells and mammalian cell lines. The mechanisms involved are unknown, but it has been proposed that PhiC31 attachment sites in the host genome recombine leading to chromosomal translocations. Results: We have studied possible effects of the PhiC31 integrase expression in human adult fibroblasts by karyotyping. All control cells were cytogenetically normal, whereas cells expressing PhiC31 integrase show chromosomal abnormalities confirming our previous results using primary... (More)
- Background: PhiC31 integrase facilitates efficient integration of transgenes into human and mouse genomes and is considered for clinical gene therapy. However recent studies have shown that the enzyme can induce various chromosomal abnormalities in primary human embryonic cells and mammalian cell lines. The mechanisms involved are unknown, but it has been proposed that PhiC31 attachment sites in the host genome recombine leading to chromosomal translocations. Results: We have studied possible effects of the PhiC31 integrase expression in human adult fibroblasts by karyotyping. All control cells were cytogenetically normal, whereas cells expressing PhiC31 integrase show chromosomal abnormalities confirming our previous results using primary embryonic fibroblasts. In order to study the early mechanisms involved we measured H2AX phosphorylation-a primary event in the response to DNA double-strand-breaks. Transient transfection with PhiC31 integrase encoding plasmids lead to an elevated number of cells positive for H2AX phosphorylation detected by immunofluorescence. Western blot analysis confirmed the upregulated H2AX phosphorylation, whereas markers for apoptosis as well as p53 and p21 were not induced. Cells transfected with plasmids encoding the Sleeping Beauty transposase remained cytogenetically normal, and in these cells less upregulation of H2AX phosphorylation could be detected. Conclusion: In primary human fibroblasts expression of PhiC31 integrase leads to a DNA damage response and chromosomal aberrations. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1444091
- author
- Liu, Jian LU ; Skjorringe, Tina ; Gjetting, Torben and Jensen, Thomas G.
- organization
- publishing date
- 2009
- type
- Contribution to journal
- publication status
- published
- subject
- in
- BMC Biotechnology
- volume
- 9
- article number
- 31
- publisher
- BioMed Central (BMC)
- external identifiers
-
- wos:000266607100002
- scopus:65849090971
- pmid:19341467
- ISSN
- 1472-6750
- DOI
- 10.1186/1472-6750-9-31
- language
- English
- LU publication?
- yes
- id
- 9eba8e46-7fcd-4f41-b279-8a0fced41ba5 (old id 1444091)
- date added to LUP
- 2016-04-01 13:05:35
- date last changed
- 2022-04-06 02:33:57
@article{9eba8e46-7fcd-4f41-b279-8a0fced41ba5,
abstract = {{Background: PhiC31 integrase facilitates efficient integration of transgenes into human and mouse genomes and is considered for clinical gene therapy. However recent studies have shown that the enzyme can induce various chromosomal abnormalities in primary human embryonic cells and mammalian cell lines. The mechanisms involved are unknown, but it has been proposed that PhiC31 attachment sites in the host genome recombine leading to chromosomal translocations. Results: We have studied possible effects of the PhiC31 integrase expression in human adult fibroblasts by karyotyping. All control cells were cytogenetically normal, whereas cells expressing PhiC31 integrase show chromosomal abnormalities confirming our previous results using primary embryonic fibroblasts. In order to study the early mechanisms involved we measured H2AX phosphorylation-a primary event in the response to DNA double-strand-breaks. Transient transfection with PhiC31 integrase encoding plasmids lead to an elevated number of cells positive for H2AX phosphorylation detected by immunofluorescence. Western blot analysis confirmed the upregulated H2AX phosphorylation, whereas markers for apoptosis as well as p53 and p21 were not induced. Cells transfected with plasmids encoding the Sleeping Beauty transposase remained cytogenetically normal, and in these cells less upregulation of H2AX phosphorylation could be detected. Conclusion: In primary human fibroblasts expression of PhiC31 integrase leads to a DNA damage response and chromosomal aberrations.}},
author = {{Liu, Jian and Skjorringe, Tina and Gjetting, Torben and Jensen, Thomas G.}},
issn = {{1472-6750}},
language = {{eng}},
publisher = {{BioMed Central (BMC)}},
series = {{BMC Biotechnology}},
title = {{PhiC31 integrase induces a DNA damage response and chromosomal rearrangements in human adult fibroblasts}},
url = {{http://dx.doi.org/10.1186/1472-6750-9-31}},
doi = {{10.1186/1472-6750-9-31}},
volume = {{9}},
year = {{2009}},
}