Tammur Jaana, Sibul Hiljar, Ustav Ene, Ustav Mart, Metspalu Andres
Institute of Molecular and Cell Biology, Tartu University, Estonian Biocentre, 23 Riia St, 51010 Tartu, Estonia.
BMC Mol Biol. 2002 Apr 19;3:5. doi: 10.1186/1471-2199-3-5.
The rationale of using bovine papillomavirus-1 (BPV-1) derived vectors in gene therapy protocols lies in their episomal maintenance at intermediate to high copy number, and stable, high-level expression of the gene products. We constructed the BPV-1 based vector harbouring the human low-density lipoprotein receptor (LDLR) gene cDNA and tested its ability to restore the function of the LDLR in the receptor-deficient cell line CHO-ldlA7.
The introduced vector p3.7LDL produced functionally active LDL receptors in the receptor-deficient cell line CHO-ldlA7 during the 32-week period of observation as determined by the internalisation assay with the labelled LDL particles.
Bovine papillomavirus type-1 (BPV-1)-derived vectors could be suitable for gene therapy due to their episomal maintenance at intermediate to high copy number and stable, high-level expression of the gene products. The constructed BPV-1 based vector p3.7LDL produced functionally active LDL receptors in the LDLR-deficient cell line CHO-ldlA7 during the 32-week period of observation. In vivo experiments should reveal, whether 1-5% transfection efficiency obtained in the current work is sufficient to bring about detectable and clinically significant lowering of the amount of circulating LDL cholesterol particles.
在基因治疗方案中使用牛乳头瘤病毒1型（BPV-1）衍生载体的基本原理在于它们能以中等至高拷贝数进行游离型维持，并稳定、高水平表达基因产物。我们构建了携带人低密度脂蛋白受体（LDLR）基因cDNA的基于BPV-1的载体，并测试了其在受体缺陷细胞系CHO-ldlA7中恢复LDLR功能的能力。
通过用标记的LDL颗粒进行内化试验确定，在32周的观察期内，导入的载体p3.7LDL在受体缺陷细胞系CHO-ldlA7中产生了具有功能活性的LDL受体。
牛乳头瘤病毒1型（BPV-1）衍生载体因其能以中等至高拷贝数进行游离型维持，并稳定、高水平表达基因产物，可能适用于基因治疗。在32周的观察期内，构建的基于BPV-1的载体p3.7LDL在LDLR缺陷细胞系CHO-ldlA7中产生了具有功能活性的LDL受体。体内实验应揭示，当前工作中获得的1%-5%的转染效率是否足以使循环LDL胆固醇颗粒的量出现可检测到的、临床上显著的降低。
