Design of a muscle cell-specific expression vector utilising human vascular smooth muscle α-actin regulatory elements

Mc, Keogh; Chen, D; Jf, Schmitt; Dennehy, Ulla; Vv, Kakkar; Nr, Lemoine

doi:10.1038/sj.gt.3300866

Cited by 22 publications

(23 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…33 The two main expression plasmids utilised in this study are the pCI/h999 expression vector ( Figure 1b) and its parent pCI (Promega), both of which have been described previously. 39 The inclusion of human VSMC ␣-actin regulatory elements in the pCI/h999 expression vector (as opposed to the CMV-containing pCI) allows the specific-targeting of VSMCs in vitro and we predict in vivo. The pCI/h999 vector has been tested in vitro in cells from a number of lineages including muscle (skeletal, cardiac and vascular smooth), endothelium (HUVEC, BAEC, PAEC), hepatocytes, fibroblasts and T cells.…”

mentioning

confidence: 85%

“…13,33 An additional myb-engrailed chimera (MybEn) and a novel myc-engrailed chimera (MycEn) were created and cloned in the expression vectors pCI and pCI/h999 (Figure 1b). 39 The sequence of the MybEn and MycEn chimeras was confirmed by DNA sequencing (data not shown). Following transfection into mammalian cells it is expected that the pCI vector will support expression of genes linked in cis in all cell types, while the genes will be expressed only in muscle cells when the pCI/h999 vector is used.…”

mentioning

confidence: 87%

“…For this reason the chimeric genes were cloned into the pCI/h999 expression vector which specifically supports transcription in muscle lineages only (Figure 2a). 39 Primary rat and rabbit VSMCs, the HUVEC cell line ECV304 or bovine aortic endothelial cells (BAECs) were then transiently transfected with pCI, pCINeo MEnT, or the chimeric proteins MybEn or MycEN in pCI or pCI/h999 as indicated. The proliferation of each population was determined by tritiated thymidine incorporation after 48 h and the level of apoptosis by TUNEL at 24 h. Transfection of the luciferase reporter gene in pCI or pCI/h999 was performed to demonstrate efficient transfection of the lineages and the specificity of the pCI/h999 vector in each case (Figure 5a).…”

Section: Unincorporated Label Was Removed By Pbs Washing and The Cementioning

confidence: 99%

“…Following transfection into mammalian cells it is expected that the pCI vector will support expression of genes linked in cis in all cell types, while the genes will be expressed only in muscle cells when the pCI/h999 vector is used. 39 To confirm this expression pattern, the MycEn and MybEn constructs in the pCI or pCI/h999 vectors were transfected into rabbit VSMCs or the HUVEC cell line ECV304 as described 39,40 and total RNA harvested 36 h later for Northern analysis. The resulting membranes were probed for engrailed or the endogenous ribosomal pro- …”

mentioning

confidence: 99%

See 3 more Smart Citations

Tissue-selective expression of dominant-negative proteins for the regulation of vascular smooth muscle cell proliferation

Dennehy

et al. 1999

Gene Ther

Self Cite

View full text Add to dashboard Cite

mentioning

confidence: 85%

mentioning

confidence: 87%

Section: Unincorporated Label Was Removed By Pbs Washing and The Cementioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Tissue-selective expression of dominant-negative proteins for the regulation of vascular smooth muscle cell proliferation

Dennehy

et al. 1999

Gene Ther

Self Cite

View full text Add to dashboard Cite

“…Natural (creatine kinase promoter-MCK, desmin) and synthetic (a-myosin heavy chain enhancer-/MCK enhancer-promoter (MHCK7)) promoters have been included in viral and nonviral vectors to achieve efficient and specific muscle expression. 87 Pompe disease, an autosomal recessive disorder caused by deficiency of acid a-glucosidase (GAA) is an important target of muscle-specific vectors. In these patients, glycogen accumulation in the lysosomes of muscle cells (cardiac and skeletal) produce severe myopathy and cardiomyopathy.…”

Section: Livermentioning

confidence: 99%

Physiological and tissue-specific vectors for treatment of inherited diseases

Toscano

Romero

Muñoz³

et al. 2010

Gene Ther

View full text Add to dashboard Cite

After more than 1500 gene therapy clinical trials in the past two decades, the overall conclusion is that for gene therapy (GT) to be successful, the vector systems must still be improved in terms of delivery, expression and safety. The recent development of more efficient and stable vector systems has created great expectations for the future of GT. Impressive results were obtained in three primary immunodeficiencies and other inherited diseases such as congenital blindness, adrenoleukodystrophy or junctional epidermolysis bullosa. However, the development of leukemia in five children included in the GT clinical trials for X-linked severe combined immunodeficiency and the silencing of the therapeutic gene in the chronic granulomatous disease clearly showed the importance of improving safety and efficiency. In this review, we focus on the main strategies available to achieve physiological or tissue-specific expression of therapeutic transgenes and discuss the importance of controlling transgene expression to improve safety. We propose that tissue-specific and/or physiological viral vectors offer the best balance between efficiency and safety and will be the tools of choice for future clinical trials in GT of inherited diseases.

show abstract

Biocompatibility of Cardiovascular Gene Delivery Catheters with Adenovirus Vectors: An Important Determinant of the Efficiency of Cardiovascular Gene Transfer

et al. 2000

View full text Add to dashboard Cite

Gene therapy approaches hold promise for the treatment of a wide variety of cardiovascular diseases. Many strategies for cardiovascular gene therapy involve catheter-mediated vector delivery via intramyocardial injection, intracoronary infusion, or direct gene transfer into the vessel wall. Several different gene delivery catheters have been developed and utilized in preclinical and clinical studies of cardiovascular gene therapy. However, rigorous studies of the biocompatibility of these catheters with gene therapy vectors have not yet been reported. In this report, we have examined the compatibility of cardiovascular gene therapy catheters and catheter constituents with first-generation E1/E3-deleted adenovirus vectors. We show that (i) currently available catheters rapidly and efficiently inactivate adenovirus vector infectivity; (ii) this inactivation is mediated by a variety of commonly used catheter constituents including stainless steel, nitinol, and polycarbonate; (iii) catheter-mediated inactivation of adenovirus vectors can be prevented by preflushing catheters with solutions of serum albumin; and (iv) it is possible to identify a set of catheter materials that are compatible with current adenovirus vectors. These results underscore the importance of catheter/vector compatibility and suggest methods for increasing the efficiency of catheter-mediated cardiovascular gene therapy.

show abstract

Design of a muscle cell-specific expression vector utilising human vascular smooth muscle α-actin regulatory elements

Cited by 22 publications

References 49 publications

Tissue-selective expression of dominant-negative proteins for the regulation of vascular smooth muscle cell proliferation

Tissue-selective expression of dominant-negative proteins for the regulation of vascular smooth muscle cell proliferation

Physiological and tissue-specific vectors for treatment of inherited diseases

Biocompatibility of Cardiovascular Gene Delivery Catheters with Adenovirus Vectors: An Important Determinant of the Efficiency of Cardiovascular Gene Transfer

Contact Info

Product

Resources

About