Showing papers by "Dieter Klein published in 1997"
TL;DR: It could be demonstrated that GFP is gener-ally superior to β-gal in terms of sensitivity, speed and noninvasiveness of assay, allowing easy direct FACS sorting of populations of transduced cells.
Abstract: Various methods for determining the expression of the β-galactosidase (β-gal) gene after retroviral transduction were compared as a means to assess retroviral titre. To allow better comparison, different retroviral vectors were constructed carrying two mutants of the green fluorescent protein and assessed as sensitive markers of retroviral gene transfer. It could be demonstrated that GFP is gener-ally superior to β-gal in terms of sensitivity, speed and noninvasiveness of assay, allowing easy direct FACS sorting of populations of transduced cells. This opens the possibility of enrichment by sorting of ex vivo transduced cells in gene therapy protocols.
59 citations
TL;DR: This system demonstrates the potential long-term therapeutic use of WAF-1/CIP- 1/SDI-1 delivered by retroviral vectors for inhibiting the growth of rapidly proliferating cells.
Abstract: Constitutive, high-level expression of the potentially therapeutic WAF-1/CIP-1/SDI-1 gene is incompatible with cell growth. A promoter conversion retroviral vector carrying the WAF-1/CIP-1/SDI-1 gene under the transcriptional control of the glucocorticoid inducible promoter of mouse mammary tumor virus was used to infect human bladder carcinoma or feline kidney cells. Reduced cell growth due to a greater proportion of cells being in the G0/G1 phase of the cell cycle was observed when WAF-1/CIP-1/SDI-1 expression was activated by addition of glucocorticoid hormone. This system demonstrates the potential long-term therapeutic use of WAF-1/CIP-1/SDI-1 delivered by retroviral vectors for inhibiting the growth of rapidly proliferating cells. Moreover, the conditional expression of genes such as WAF-1/CIP-1/SDI-1 from such retroviral vectors may facilitate analysis of their function.
17 citations