Optimization of transgene expression at the posttranscriptional level in neural cells: Implications for gene therapy

被引:40
作者
Brun, S [1 ]
Faucon-Biguet, N [1 ]
Mallet, J [1 ]
机构
[1] Hop La Pitie Salpetriere, Lab Genet Mol Neurotransmiss & Proc Neurodegenera, CNRS, UMR 7091, F-75013 Paris, France
关键词
WPRE; tau 3 ' UTR; TH 3 ' UTR; APP 5 ' UTR; gene expression; posttranscriptional enhancer; neural cells; lentiviral vector;
D O I
10.1016/S1525-0016(03)00097-2
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Gene delivery vectors need to fulfill several efficacy and safety criteria before they can be used in humans. Successful clinical application requires effective transgene expression with a minimum of vector-associated toxicity. We describe the use of posttranscriptional regulatory elements in plasmid and lentiviral vectors coding for luciferase. These constructs allow high-level gene expression in both neuronal and glial cells. Of the several elements that we tested, WPRE gave the highest level of expression. Further enhancements were obtained when WPRE was combined with sequences corresponding to the 3' or 5' untranslated regions (UTR) of eukaryotic mRNAs (tau 3'UTR, TH 3'UTR, and APP 5'UTR). In neuronal cells, WPRE and both tau 3'UTR and APP 5'UTR had an additive effect on expression. The combination of the three elements increased the basal level of expression by up to 26-fold. In glial cells, WPRE and APP 5'UTR had additive effects on expression, and their combination increased expression up to 10-fold. These results provide important information regarding the development of optimal CNS gene transfer vectors not only for gene therapy but also for the study of gene function.
引用
收藏
页码:782 / 789
页数:8
相关论文
共 36 条
[1]   TISSUE-SPECIFIC EXPRESSION AND CDNA STRUCTURE OF A HUMAN TRANSCRIPT ENCODING A NUCLEIC-ACID BINDING [OLIGO(DC)] PROTEIN RELATED TO THE PRE-MESSENGER-RNA BINDING-PROTEIN-K [J].
AASHEIM, HC ;
LOUKIANOVA, T ;
DEGGERDAL, A ;
SMELAND, EB .
NUCLEIC ACIDS RESEARCH, 1994, 22 (06) :959-964
[2]   Efficient 3′-end formation of human β-globin mRNA in vivo requires sequences within the last intron but occurs independently of the splicing reaction [J].
Antoniou, M ;
Geraghty, F ;
Hurst, J ;
Grosveld, F .
NUCLEIC ACIDS RESEARCH, 1998, 26 (03) :721-729
[3]  
ARANDAABREU GE, 1999, J MOL NEUROSCI, V12, P1
[4]   Identification of 3′UTR region implicated in tau mRNA stabilization in neuronal cells [J].
Aronov, S ;
Marx, R ;
Ginzburg, L .
JOURNAL OF MOLECULAR NEUROSCIENCE, 1999, 12 (02) :131-145
[5]   COMPARISON OF INTRON-DEPENDENT AND INTRON-INDEPENDENT GENE-EXPRESSION [J].
BUCHMAN, AR ;
BERG, P .
MOLECULAR AND CELLULAR BIOLOGY, 1988, 8 (10) :4395-4405
[6]   A GENERIC INTRON INCREASES GENE-EXPRESSION IN TRANSGENIC MICE [J].
CHOI, T ;
HUANG, M ;
GORMAN, C ;
JAENISCH, R .
MOLECULAR AND CELLULAR BIOLOGY, 1991, 11 (06) :3070-3074
[7]   Woodchuck hepatitis virus contains a tripartite posttranscriptional regulatory element [J].
Donello, JE ;
Loeb, JE ;
Hope, TJ .
JOURNAL OF VIROLOGY, 1998, 72 (06) :5085-5092
[8]   COMPLETE CODING SEQUENCE OF RAT TYROSINE-HYDROXYLASE MESSENGER-RNA [J].
GRIMA, B ;
LAMOUROUX, A ;
BLANOT, F ;
BIGUET, NF ;
MALLET, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1985, 82 (02) :617-621
[9]  
HAMPRECHT B, 1985, METHOD ENZYMOL, V109, P341
[10]   Four highly stable eukaryotic mRNAs assemble 3' untranslated region RNA-protein complexes sharing cis and trans components [J].
Holcik, M ;
Liebhaber, SA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (06) :2410-2414