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| Expression of a Chaining miR-shRNA for Simultaneous Knockdown of srGAP Family Members |
| MA Yue, DAI Yun-Kai, CHEN Keng, MI Ya-Jing, JIN Wei-Lin |
| (School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai 200240, China) |
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Abstract To resolve the partial compensation effect of the different srGAPs in neuronal development, a miR-shRNA expression system was constructed which can down regulate srGAP family members srGAP1—3 simultaneously. Multiple miRshRNA oligos targeting different srGAPs were designed, synthesized and then inserted into the miRNA expression vector to generate miRshRNA against srGAP1—3 respectively. Western blot was performed to detect the knockdown effect of different miR-shRNAs, the three miRshRNA with the most obvious knockdown effect to srGAP1—3 respectively were selected. Then the miR-shRNAs were chained together in one vector to generate a chaining miRshRNA expression vector. The results show that the effective miRshRNA against corresponding srGAP1—3 are generated and named J17, J24 and J33. Furthermore, the chaining miR-shRNA J1+2+3 against srGAP1—3 is obtained.
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Received: 20 March 2011
Published: 28 May 2012
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