Abstract
Cardiomyocytes are quite resistant to gene transfer using standard techniques. We developed an expression vector carrying an attenuated but infectious and replicative coxsackievirus B3 (CVB3) genome, and unique ClaI-StuI cloning sites for an exogenous gene, whose product can be released from the nascent viral polyprotein by 2Apro cleavage. This vector was tested as an expression vehicle for green fluorescent protein (GFP). The vector transiently expressed GFP in cell cultures for at least ten passages and delivered functional GFP to the infected cardiomyocytes for at least 6 days. Moreover, the recombinant viruses showed virulence attenuation in vitro and in vivo. The findings suggest that the recombinant CVB3 vector could be a useful tool for viral tracking study and delivering exogenous proteins to cardiomyocytes.
Highlights
Gene therapy is a promising treatment of choice for myocardial diseases [1,2,3]; cardiomyocytes are quite resistant to gene transfer by standard techniques [4], and developing an efficient gene delivery method remains a major challenge for cardiac gene therapy
Lentiviruses can replicate in nondividing cells and are considered as potential viral vectors for cardiac gene therapy [5], these viruses may cause concern because they are derived from human immunodeficiency virus type 1 (HIV1)
We have developed an efficient method to express an exogenous protein in cardiomyocytes in vivo using an attenuated Coxsackievirus B3 (CVB3) as a gene delivery vector
Summary
Due to high expression efficiency and ability to transfect a wide range of cell types, recombinant viral vectors are commonly used for gene transfer. Coxsackievirus B3 (CVB3) has a significant advantage over other viral vectors by replicate in nondividing cells, such as myocytes. Lentiviruses can replicate in nondividing cells and are considered as potential viral vectors for cardiac gene therapy [5], these viruses may cause concern because they are derived from human immunodeficiency virus type 1 (HIV1). Coxsackievirusadenovirus receptor (CAR) and decay accelerating factor (CD55) have been shown to enable coxsackievirus attach and enter the cell [6]. CVB3 could be exploited as a vector to deliver exogenous genes to the heart or as a viral tracking vector
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