Difference between revisions of "Part:BBa K2208001:Experience"
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| + | ===Applications of BBa_K2208001=== | ||
| − | + | ====Usage and Biology==== | |
| − | + | tPep is a white fat tissue targeting peptide. To enhance the accuracy of siRNA delivery system and improve targeting index of drugs, tPep was displayed on the surface of the exosomes containing our previously designed siRNAs, allowing us to target recipient cells in vivo efficiently. | |
| − | + | Lamp-2b is a protein found specifically abundant in exosomal membranes. So we connect tPep with Lamp2b by a glycine-linker, and promote the expression using cmv promoter. In that way, the tPep exosomes (tPep-Exos) are endowed with site-specific identification ability and were purified from cell culture supernatants and loaded with siRNA. | |
| − | === | + | ====Characterization==== |
| + | To prove the targeting property of tPep exosomes, we co-incubated them respectively with 3T3L1 cells (mouse adipose tissues) and C2C12 (mouse myoblasts). Also we set parallel experiments using exosomes without tPep-lamp 2b fusion protein. It was thrilling to see only tPep exosomes accumulated in the 3T3L1 cells while the other groups’ results were contrary. This fully illustrates tPep exosomes can target precisely to the adipose tissues. | ||
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| + | [[File:NJU_China_iGEM_2017_test0000.jpeg|200px]] | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 14:16, 27 October 2017
Applications of BBa_K2208001
Usage and Biology
tPep is a white fat tissue targeting peptide. To enhance the accuracy of siRNA delivery system and improve targeting index of drugs, tPep was displayed on the surface of the exosomes containing our previously designed siRNAs, allowing us to target recipient cells in vivo efficiently. Lamp-2b is a protein found specifically abundant in exosomal membranes. So we connect tPep with Lamp2b by a glycine-linker, and promote the expression using cmv promoter. In that way, the tPep exosomes (tPep-Exos) are endowed with site-specific identification ability and were purified from cell culture supernatants and loaded with siRNA.
Characterization
To prove the targeting property of tPep exosomes, we co-incubated them respectively with 3T3L1 cells (mouse adipose tissues) and C2C12 (mouse myoblasts). Also we set parallel experiments using exosomes without tPep-lamp 2b fusion protein. It was thrilling to see only tPep exosomes accumulated in the 3T3L1 cells while the other groups’ results were contrary. This fully illustrates tPep exosomes can target precisely to the adipose tissues.
User Reviews
UNIQ9af76f9ee4da5095-partinfo-00000000-QINU UNIQ9af76f9ee4da5095-partinfo-00000001-QINU