| Autor: |
Alami R; Department of Medicine, Albert Einstein College of Medicine, Bronx, New York 10461, USA., Gilman JG, Feng YQ, Marmorato A, Rochlin I, Suzuka SM, Fabry ME, Nagel RL, Bouhassira EE |
| Jazyk: |
angličtina |
| Zdroj: |
Blood cells, molecules & diseases [Blood Cells Mol Dis] 1999 Apr; Vol. 25 (2), pp. 110-9. |
| DOI: |
10.1006/bcmd.1999.0235 |
| Abstrakt: |
Our current strategy for gene therapy of sickle cell anemia involves retroviral vectors capable of transducing "designer" globin genes that code for novel anti-sickling globins (while resisting digestion by a ribozyme), coupled with the expression of a hammerhead ribozyme that can selectively cleave the human beta s mRNA. In this report, we have tested in vivo an anti-beta s hammerhead ribozyme embedded within a cDNA coding for the luciferase reporter gene driven by the human beta-globin promoter and hyper-sensitive sites 3 and 4 of the locus control region. We have created mice transgenic for this luciferase-ribozyme construct and bred the ribozyme transgene into mice that were already transgenic for the human beta s gene. We then measured expression of the beta s transgene at the protein and RNA levels by HPLC and primer extension. The presence of the ribozyme was associated with a statistically significant reduction in the level of beta s mRNA in spleen stress reticulocytes (from 60.5 +/- 4.1% to 52.9 +/- 4.2%) and in the percentage of beta s globin chains in very young mice (from 44.5 +/- 0.6% to 40.8 +/- 0.7%). These results demonstrate that it is possible to decrease the concentration of beta s chains and mRNA with the help of a hammerhead ribozyme. While the enormous amount of globin mRNA in reticulocytes is a challenge for ribozyme technology, the exquisite dependence of the delay time for formation of Hb S nuclei on the concentration of Hb S in red blood cells suggests that even a modest reduction in Hb S concentration would have therapeutic value. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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