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  • Title: Gene therapy for streptozotocin-induced diabetic mice by electroporational transfer of naked human insulin precursor DNA into skeletal muscle in vivo.
    Author: Yin D, Tang JG.
    Journal: FEBS Lett; 2001 Apr 20; 495(1-2):16-20. PubMed ID: 11322940.
    Abstract:
    Transfer of naked plasmid with insulin precursor DNA into skeletal muscle of streptozotocin (STZ)-induced diabetic mice through electroporation and detection of gene expression is described. Four different human insulin precursor DNA fragments were inserted into pcDNA3.1(-), downstream of a CMV promoter. Three of them, with a secretion signal sequence, succeeded in lowering blood glucose at a range of 30-50% in STZ diabetic mice. The other, with a synthetic DNA fragment encoding human proinsulin, failed. The mortality rate of very seriously STZ diabetic mice was reduced significantly by the treatment. The circulating insulin-like protein (mouse insulin, human proinsulin, or intermediates during conversion of proinsulin to insulin) level in the blood of less seriously STZ diabetic mice treated with the human preproinsulin gene with an intron was about 15-23 microU/ml, while that of STZ diabetic mice treated with empty vector was only about 6 microU/ml and that of normal mice was about 18 microU/ml. Transcription of the three human insulin precursor DNAs in mouse skeletal muscle was also detected by RT-PCR. The human preproinsulin gene with the intron showed a slightly higher potency in reducing blood glucose of mildly diabetic mice. These studies indicate that the skeletal muscle transferred with appropriate preproinsulin DNA by electroporation in vivo can secrete insulin-like protein resulting in reduction of blood glucose, and a basal blood insulin level can be achieved for at least 1 month.
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