Generation of a new Gateway-compatible inducible lentiviral vector platform allowing easy derivation of co-transduced cells

被引：12

作者：

De Groote, Philippe ^{[1
,2
,3
]}

Grootjans, Sasker ^{[1
,2
,3
]}

Lippens, Saskia ^{[1
,2
,3
]}

Eichperger, Chantal ^{[1
,2
,3
]}

Leurs, Kirsten ^{[1
,2
,3
]}

Kahr, Irene ^{[1
,2
,3
]}

Tanghe, Giel ^{[1
,2
,3
]}

Bruggeman, Inge ^{[1
,2
,3
]}

De Schamphelaire, Wouter

Urwyler, Corinne ^{[1
,2
,3
]}

Vandenabeele, Peter ^{[1
,2
,3
]}

Haustraete, Jurgen ^{[1
,2
,3
]}

Declercq, Wim ^{[1
,2
,3
]}

机构：

[1] Flanders Inst Biotechnol VIB, Inflammat Res Ctr, B-9052 Ghent, Belgium

[2] Univ Ghent, Dept Biomed Mol Biol, B-9000 Ghent, Belgium

[3] Univ Ghent, BCCM LMBP Plasmid Collect, Dept Biomed Mol Biol, B-9000 Ghent, Belgium

来源：

BIOTECHNIQUES | 2016年 / 60卷 / 05期

关键词：

lentiviral vectors; co-inducible expression; Tet-On; Gateway; NF-KAPPA-B; MAMMALIAN-CELLS; RNA INTERFERENCE; RECOMBINANT PROTEINS; GENE-EXPRESSION; IN-VIVO; SYSTEM; KINASE; LINE;

D O I：

10.2144/000114417

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

In contrast to most common gene delivery techniques, lentiviral vectors allow targeting of almost any mammalian cell type, even non-dividing cells, and they stably integrate in the genome. Therefore, these vectors are a very powerful tool for biomedical research. Here we report the generation of a versatile new set of 22 lentiviral vectors with broad applicability in multiple research areas. In contrast to previous systems, our platform provides a choice between constitutive and/or conditional expression and six different C-terminal fusions. Furthermore, two compatible selection markers enable the easy derivation of stable cell lines co-expressing differently tagged transgenes in a constitutive or inducible manner. We show that all of the vector features are functional and that they contribute to transgene overexpression in proof-of-principle experiments.

引用

页码：252 / 259

页数：8

共 21 条

[1] Daedalus: a robust, turnkey platform for rapid production of decigram quantities of active recombinant proteins in human cell lines using novel lentiviral vectors [J].