Transgene stability for three replication-competent murine leukemia virus vectors
Research output: Contribution to journal › Journal article › Research › peer-review
Standard
Transgene stability for three replication-competent murine leukemia virus vectors. / Duch, Mogens R.; Carrasco, Maria L; Jespersen, Thomas; Hansen, Bettina Dencker; Pedersen, Finn Skou; Jespersen, Thomas.
In: Gene, Vol. 329, 31.03.2004, p. 61-9.Research output: Contribution to journal › Journal article › Research › peer-review
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - Transgene stability for three replication-competent murine leukemia virus vectors
AU - Duch, Mogens R.
AU - Carrasco, Maria L
AU - Jespersen, Thomas
AU - Hansen, Bettina Dencker
AU - Pedersen, Finn Skou
AU - Jespersen, Thomas
PY - 2004/3/31
Y1 - 2004/3/31
N2 - Retroviral vectors that are able to sustain multiple rounds of replication may find many applications. However, one critical feature of such vectors is the ability to maintain an intact transgene cassette during repeated rounds of replication. We here report on the stability of a translational cassette consisting of an internal ribosome entry site followed by the enhanced green fluorescent protein coding sequence inserted in different configurations into murine leukemia virus genomes. In two of the constructs, the insert was located in the upstream part of the U3 region while in the third construct it was inserted in the 3' untranslated region of the viral genome. Furthermore, in two of the constructs, the translational cassette was flanked by a direct repeat, while no such structure flanked the third construct. Our results show that deletion of the heterologous translational cassette is observed for all constructs upon extended cell culture and that the number of replication rounds before revertants are detected can be postponed by decreasing the length of the repeat flanking the translational cassette.
AB - Retroviral vectors that are able to sustain multiple rounds of replication may find many applications. However, one critical feature of such vectors is the ability to maintain an intact transgene cassette during repeated rounds of replication. We here report on the stability of a translational cassette consisting of an internal ribosome entry site followed by the enhanced green fluorescent protein coding sequence inserted in different configurations into murine leukemia virus genomes. In two of the constructs, the insert was located in the upstream part of the U3 region while in the third construct it was inserted in the 3' untranslated region of the viral genome. Furthermore, in two of the constructs, the translational cassette was flanked by a direct repeat, while no such structure flanked the third construct. Our results show that deletion of the heterologous translational cassette is observed for all constructs upon extended cell culture and that the number of replication rounds before revertants are detected can be postponed by decreasing the length of the repeat flanking the translational cassette.
KW - 3T3 Cells
KW - Animals
KW - Base Sequence
KW - Cell Line
KW - DNA, Recombinant
KW - Flow Cytometry
KW - Gene Expression
KW - Genetic Vectors
KW - Green Fluorescent Proteins
KW - Humans
KW - Leukemia Virus, Murine
KW - Luminescent Proteins
KW - Mice
KW - Molecular Sequence Data
KW - Transfection
KW - Transgenes
KW - Virus Replication
U2 - 10.1016/j.gene.2003.12.032
DO - 10.1016/j.gene.2003.12.032
M3 - Journal article
C2 - 15033529
VL - 329
SP - 61
EP - 69
JO - Gene
JF - Gene
SN - 0378-1119
ER -
ID: 33017346