Difference between revisions of "Part:BBa K2208001"
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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. The adipocyte targeting capability of exosomes was conferred by engineering the human embryonic kidney 293 (HEK293) cells to express lysosome-associated membrane glycoprotein 2b (Lamp2b), a well-characterized exosomal membrane protein, fused with tPep (CKGGRAKDC) for integrin. 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. | 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. The adipocyte targeting capability of exosomes was conferred by engineering the human embryonic kidney 293 (HEK293) cells to express lysosome-associated membrane glycoprotein 2b (Lamp2b), a well-characterized exosomal membrane protein, fused with tPep (CKGGRAKDC) for integrin. 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. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
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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. | 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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Latest revision as of 17:54, 25 October 2017
A coding sequence of tPep-Lamp2b fusion protein and position it outside the membrane.
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. The adipocyte targeting capability of exosomes was conferred by engineering the human embryonic kidney 293 (HEK293) cells to express lysosome-associated membrane glycoprotein 2b (Lamp2b), a well-characterized exosomal membrane protein, fused with tPep (CKGGRAKDC) for integrin. 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.
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.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1276
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 199