Functional Analysis of Drosophila HSP70 Promoter with Different HSE Numbers in Human Cells |
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| Authors: | Nadezda Kust Ekaterina Rybalkina Ilya Mertsalov Ekaterina Savchenko Alexander Revishchin Gali Pavlova |
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| Institution: | 1. Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia.; 2. Institute of Medicine and Cell Transplantation, Moscow, Russia.; 3. Ltd Apto-pharm, Moscow, Russia.; University of Arkansas for Medical Sciences, United States of America, |
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| Abstract: | The activation of genetic constructs including the Drosophila hsp70 promoter with four and eight HSE sequences in the regulatory region has been described in human cells. The promoter was shown to be induced at lower temperatures compared to the human hsp70 promoter. The promoter activity increased after a 60-min heat shock already at 38°C in human cells. The promoter activation was observed 24 h after heat shock for the constructs with eight HSEs, while those with four HSEs required 48 h. After transplantation of in vitro heat-shocked transfected cells, the promoter activity could be maintained for 3 days with a gradual decline. The promoter activation was confirmed in vivo without preliminary heat shock in mouse ischemic brain foci. Controlled expression of the Gdnf gene under a Drosophila hsp70 promoter was demonstrated. This promoter with four and eight HSE sequences in the regulatory region can be proposed as a regulated promoter in genetic therapeutic systems. |
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