Efficient gene transfer in skeletal muscle with AAV-derived bicistronic vector using the FGF-1 IRES

Delluc-Clavieres, A.; Bec, Christine Le; Berghe, L. Van Den; Conte, Caroline; Allo, Valérie; Danos, Olivier; Prats, Anne-Catherine

doi:10.1038/gt.2008.49

Cited by 13 publications

(13 citation statements)

References 34 publications

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“…In vivo imaging was performed with a cooled CCD camera (1394 ORCA II, Hamamatsu Photonics) using mice transduced with a bicistronic AAV vector expressing the Firefly luciferase under control of FGF1 IRES A, as previously described (26). Eighteen days after transduction, muscles were cardiotoxin-treated and imaged 3 and 7 days later.…”

Section: Methodsmentioning

confidence: 99%

Fibroblast growth factor 1 induced during myogenesis by a transcription–translation coupling mechanism

Conte

Ainaoui

Delluc-Clavieres

et al. 2009

Nucleic Acids Research

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Fibroblast growth factor 1 (FGF1) is involved in muscle development and regeneration. The FGF1 gene contains four tissue-specific promoters allowing synthesis of four transcripts with distinct leader regions. Two of these transcripts contain internal ribosome entry sites (IRESs), which are RNA elements allowing mRNA translation to occur in conditions of blockade of the classical cap-dependent mechanism. Here, we investigated the function and the regulation of FGF1 during muscle differentiation and regeneration. Our data show that FGF1 protein expression is induced in differentiating myoblasts and regenerating mouse muscle, whereas siRNA knock-down demonstrated FGF1 requirement for myoblast differentiation. FGF1 induction occurred at both transcriptional and translational levels, involving specific activation of both promoter A and IRES A, whereas global cap-dependent translation was inhibited. Furthermore, we identified, in the FGF1 promoter A distal region, a cis-acting element able to activate the IRES A-driven translation. These data revealed a mechanism of molecular coupling of mRNA transcription and translation, involving a unique process of IRES activation by a promoter element. The crucial role of FGF1 in myoblast differentiation provides physiological relevance to this novel mechanism. This finding also provides a new insight into the molecular mechanisms linking different levels of gene expression regulation.

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Section: Methodsmentioning

confidence: 99%

Fibroblast growth factor 1 induced during myogenesis by a transcription–translation coupling mechanism

Conte

Ainaoui

Delluc-Clavieres

et al. 2009

Nucleic Acids Research

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“…Although a gene transcribed upstream of IRES can be translated at a much higher level than the downstream gene, 22 there was evidence that the ratio of translation between each gene remained pretty much stable. 23,24 The bicistronic FM-1 lentivector in our study enables the independent expression of the BLI gene in conjunction with the fluorescent probe Venus, a variant of GFP.…”

Section: Discussionmentioning

confidence: 99%

Comparison of red-shifted firefly luciferase Ppy RE9 and conventional Luc2 as bioluminescence imaging reporter genes for <italic>in vivo</italic> imaging of stem cells

Liang

2012

J. Biomed. Opt

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Abstract. One critical issue for noninvasive imaging of transplanted bioluminescent cells is the large amount of light absorption in tissue when emission wavelengths below 600 nm are used. Luciferase with a red-shifted spectrum can potentially bypass this limitation. We assessed and compared a mutant of firefly luciferase (Ppy RE9, PRE9) against the yellow luciferase luc2 gene for use in cell transplantation studies. C17.2 neural stem cells expressing PRE9-Venus and luc2-Venus were sorted by flow cytometry and assessed for bioluminescence in vitro in culture and in vivo after transplantation into the brain of immunodeficient Rag2-/-mice. We found that the luminescence from PRE9 was stable, with a peak emission at 620 nm, shifted to the red compared to that of luc2. The emission peak for PRE9 was pH-independent, in contrast to luc2, and much less affected by tissue absorbance compared to that of luc2. However, the total emitted light radiance from PRE9 was substantially lower than that of luc2, both in vitro and in vivo. We conclude that PRE9 has favorable properties as compared to luc2 in terms of pH independence, redshifted spectrum, tissue light penetration, and signal quantification, justifying further optimization of protein expression and enzymatic activity.

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“…27,[41][42][43][44] In this study, to identify novel IRES sequences that can improve expression vectors, we conducted in vitro and in vivo screening of 13 IRES elements from viral or cellular origin.…”

Section: Discussionmentioning

confidence: 99%

“…To address this issue, several groups have sought to identify alternative cellular and viral IRES elements that initiate protein expression more efficiently than EMCV IRES. 15,[19][20][21][22][23][24][25][26] Cellular IRES elements such as eukaryotic initiation factor 4G, 15 immunoglobulin heavy chain binding protein, 15 c-myc proto-oncogene, 15 vascular endothelial growth factor 15 and fibroblast growth factor-1 27 IRES have been reported to show higher efficiency than EMCV IRES. In this study, we searched to identify an IRES(s) with higher translation efficiency than the EMCV IRES both in vitro and in vivo.…”

Section: Introductionmentioning

confidence: 99%

In vitro and in vivo comparison of viral and cellular internal ribosome entry sites for bicistronic vector expression

et al. 2011

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Bicistronic vectors are essential to achieve efficient expression of multiple genes in gene therapy protocols and biomedical applications. Internal ribosome entry site (IRES) elements have been utilized to initiate expression of an additional protein from a bicistronic vector. The IRES element commonly used in current bicistronic vectors originates from the encephalomyocarditis virus (EMCV). As IRES-mediated translation is dependent on availability of IRES trans-acting factors, which vary between cell types and species, adequate gene expression from the EMCV IRES element is not always achieved. To identify a novel IRES element that mediates gene expression consistently with a higher efficiency than the EMCV IRES, we tested 13 bicistronic reporter constructs containing different viral and cellular IRES elements. The in vitro screening in human and mouse fibroblast and hepatocarcinoma cells revealed that the vascular endothelial growth factor and type 1 collagen-inducible protein (VCIP) IRES was the only IRES element that directed translation more efficiently than the EMCV IRES in all cell lines. Furthermore, the VCIP IRES initiated greater reporter expression levels than the EMCV IRES in transfected mouse livers. These results suggest that VCIP-IRES containing vectors improve gene expression compared with those harboring an EMCV-IRES. This could increase the potential benefits of bicistronic vectors for experimental and therapeutic purposes.

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Efficient gene transfer in skeletal muscle with AAV-derived bicistronic vector using the FGF-1 IRES

Cited by 13 publications

References 34 publications

Fibroblast growth factor 1 induced during myogenesis by a transcription–translation coupling mechanism

Fibroblast growth factor 1 induced during myogenesis by a transcription–translation coupling mechanism

Comparison of red-shifted firefly luciferase Ppy RE9 and conventional Luc2 as bioluminescence imaging reporter genes for <italic>in vivo</italic> imaging of stem cells

In vitro and in vivo comparison of viral and cellular internal ribosome entry sites for bicistronic vector expression

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