Researchers at National Institute of Dental and Craniofacial Research Target Luminescent Proteins (Integration of the hybrid adenoretroviral vector AdLTR-luc involves both MoMLV elements flanking the transgene)
By a News Reporter-Staff News Editor at Health & Medicine Week -- Investigators publish new report on Proteins. According to news reporting from Bethesda, Maryland, by NewsRx journalists, research stated, "Vector delivery is still a bottleneck for gene therapy. To overcome some disadvantages of adenoviral and retroviral vectors, we developed a hybrid vector."
The news correspondents obtained a quote from the research from the National Institute of Dental and Craniofacial Research, "This hybrid vector, AdLTR-luc, was created by adding two elements from Moloney murine leukemia virus (MoMLV) flanking the luciferase cDNA into an E1/E3-deleted, replication deficient serotype 5 adenovirus vector (Zheng et al., Nature Biotechnol, 2000), and demonstrated that the MoMLV element upstream of the luciferase cDNA was broken during the integration event. The purpose of the current study was to determine if the MoMLV element downstream of the luciferase cDNA was also broken when integration occurred. We used the same A5 cell clones (#10 and 11) from the earlier the paper along with restriction endonuclease digestions, plus Southern hybridization, and PCR. Southern hybridization indicated that the luciferase cDNA was intact in the cloned cells. Results from Xho I and Sal I digestions showed that integration occurred in cloned cells. Southern hybridizations after Nco I digestion suggested that there was a break in both MoMLV elements, upstream and downstream of the luciferase cDNA. After DNA digestion with Not I, hybridization analyses indicated that the MoMLV upstream element was broken during integration. Digestion of genomic DNA with either Xba I/Kpn I, Bam HI/Sac I, or Bam HI/Nco I demonstrated that the MoMLV downstream element was also broken during integration. A PCR assay was unable to amplify the junctional region between the downstream MoMLV element and the adenoviral E2B gene, consistent with a break in that element."
According to the news reporters, the research concluded: "Although AdLTR-luc integration is atypical (Zheng et al., Nature Biotechnol, 2000), the present results suggest that both MoMLV elements have important roles in this event."
For more information on this research see: Integration of the hybrid adenoretroviral vector AdLTR-luc involves both MoMLV elements flanking the transgene. International Journal of Medical Sciences, 2014;11(8):803-9 (see also Proteins).
Our news journalists report that additional information may be obtained by contacting C. Zheng, Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892-1190, United States.
Keywords for this news article include: Bethesda, Maryland, United States, North and Central America, Enzymes and Coenzymes, Luciferases, Luminescent Proteins, Viral DNA.
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