Glucose 6-phosphate dehydrogenase expression is less prone to variegation when driven by its own promoter. (Articolo in rivista)

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  • Glucose 6-phosphate dehydrogenase expression is less prone to variegation when driven by its own promoter. (Articolo in rivista) (literal)
Anno
  • 2001-01-01T00:00:00+01:00 (literal)
Alternative label
  • De Angioletti M; Rovira A; Notaro R; Camacho Vanegas O; Sadelain M; Luzzatto L. (2001)
    Glucose 6-phosphate dehydrogenase expression is less prone to variegation when driven by its own promoter.
    in Gene (Amst.)
    (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
  • De Angioletti M; Rovira A; Notaro R; Camacho Vanegas O; Sadelain M; Luzzatto L. (literal)
Pagina inizio
  • 221 (literal)
Pagina fine
  • 231 (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume
  • 267 (literal)
Rivista
Note
  • ISI Web of Science (WOS) (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
  • Department of Human Genetics, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021, USA, Istituto Internazionale di Genetica e Biofisica (IIGB-CNR), Napoli, Italy. (literal)
Titolo
  • Glucose 6-phosphate dehydrogenase expression is less prone to variegation when driven by its own promoter. (literal)
Abstract
  • The ability to transfer permanently genes into mammalian cells makes retroviruses suitable vectors for the ultimate purpose of treating inherited genetic disease. However, expression of the retrovirally transferred genes is variable (position effect and expression variegation) because retroviruses are highly susceptible to the influence of the host genome sequences which flank the integration site. We have investigated this phenomenon with respect to the human housekeeping enzyme, glucose 6-phosphate dehydrogenase (hG6PD). We have constructed retroviral vectors in which the hG6PD cDNA is driven by either of two conventional retroviral promoters and enhancers from the Moloney Murine Leukemia Virus (MMLV) and the Myeloproliferative Sarcoma Virus (MPSV) long terminal repeats (LTR) or by the hG6PD own promoter replacing most of enhancer and promoter LTR (GRU5). We have compared the activity of retrovirally transferred hG6PD driven by these promoters after retroviral integration in bulk cultures and in individual clones of murine fibroblasts. The level of hG6PD expressed by the hG6PD promoter of GRU5-G6PD was significantly lower than that expressed by conventional retroviral vectors. However, analysis of the single copy clones showed less variation of expression with GRU5-G6PD (coefficient of variation, CV, 35.5%) than with conventional vectors (CV, 58.9%). Thus we have several vectors competent for reliable transfer and expression of hG6PD. The hG6PD promoter provides reproducible expression of hG6PD and limits the variability of expression. This decreased variability is important in order to help ensuring a consistent level of delivery of the needed gene product in future therapeutic protocols. (literal)
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