Girod A, Cosset F L, Verdier G, Ronfort C
Centre de Génétique Moléculaire et Cellulaire, CNRS UMR 106, INRA, Université Claude Bernard, Villeurbanne, France.
Virology. 1995 Jun 1;209(2):671-5. doi: 10.1006/viro.1995.1302.
We have previously described avian leukosis virus-based packaging cell lines that express gag, pol, and env proteins from two transcomplementing genomes and produce helper-free stocks of retroviral vectors with different host ranges. In this report, we demonstrated that (i) despite the deletion of the psi packaging sequence, the packaging-defective transcomplementing retroviral transcripts were packaged into virions at a level that could reach 2.3% of a wild-type virus packaging level and (ii) despite deletion of the 3' LTR, these genomes were transferred along with the vector to target cells. As these genomes were also bearing a selectable gene, titers of the resulting contaminant particles could be estimated, depending on the cell line to be between 0 and 6 infectious particles/ml of supernatant.
我们之前描述过基于禽白血病病毒的包装细胞系,这些细胞系从两个反式互补基因组表达gag、pol和env蛋白,并产生具有不同宿主范围的无辅助病毒的逆转录病毒载体储备。在本报告中,我们证明:(i)尽管缺失了ψ包装序列,但包装缺陷型反式互补逆转录病毒转录本仍能以可达野生型病毒包装水平2.3%的比例包装到病毒粒子中;(ii)尽管缺失了3' LTR,但这些基因组仍会与载体一起转移到靶细胞中。由于这些基因组也携带一个可选择基因,因此可以根据细胞系估计所得污染颗粒的滴度,上清液中每毫升的感染性颗粒数在0至6之间。