Difference between revisions of "Part:BBa K2455003"

Revision as of 22:41, 31 October 2017

Cell-Penetrating USER Cassette

This biobrick contains an AsiSI/Nb.BSMI USER Cassette with a N-terminal nona-arginine (R9) Cell-penetrating peptide which gives proteins inserted into the cassette the ability to pass through plasma membranes. In addition the USER Cassette carries a N-terminal hexa-histidine tag which allows for easy purification.

For our study we demonstrated that proteins inserted into the cassette can be purified, using the histidine-tag, and subsequently transferred into E. coli cells through the use of the R9 peptide.

In addition; previous studies have shown proteins associated with R9, either covalently or non-covalently, to be able to enter a variety of cell types (Chang et al.).

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Usage and Biology

Cell-Penetrating Peptides (CPPs) are small peptides, typically rich in basic residues, which are able to facilitate transport of a wide variety of cargoes across plasma membranes. Their origin in nature comes from viral domains such as the viral HIV tat domain (Eudes & Chugh, 2008). In recent years research have been done into making synthetic CPPs, especially peptides constructed solely from arginine residues have been of interest. The arginine rich sequence has been shown to trigger endocytosis in a wide range of cell types, including onion and potato cells (Chang et al.).

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 and subsequently transferred into <i>E. coli</i> cells through the use of the R9 peptide.
-In addition; previous studies have shown proteins associated with R9, either covalently or non-covalently, to be able to enter a variety of cell types (link!).
+In addition; previous studies have shown proteins associated with R9, either covalently or non-covalently, to be able to enter a variety of cell types ([https://academic.oup.com/pcp/article/46/3/482/1817347 Chang <i>et al.</i>]).
 <!-- Add more about the biology of this part here
@@ Line 24: / Line 24: @@
 <span class='h3bb'><font size="5"><b>Usage and Biology</b></font></span>
-Cell-Penetrating Peptides (CPPs) are small peptides, typically rich in arginines, which are able to facilitate transport of a wide variety of cargoes across plasma membranes. Their origin in nature comes from viral domains such as the viral HIV tat domain ([https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2634491/ Eudes & Chugh, 2008]). In recent years research have been done into making synthetic CPPs, especially peptides constructed solely from arginine residues have been of interest. The arginine rich sequence has been shown to trigger endocytosis in a wide range of cell types, including onion and potato cells. These experiments have shown that GFP connected to a CPP has entered the cells contained in vesicles (ref).
+Cell-Penetrating Peptides (CPPs) are small peptides, typically rich in basic residues, which are able to facilitate transport of a wide variety of cargoes across plasma membranes. Their origin in nature comes from viral domains such as the viral HIV tat domain ([https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2634491/ Eudes & Chugh, 2008]). In recent years research have been done into making synthetic CPPs, especially peptides constructed solely from arginine residues have been of interest. The arginine rich sequence has been shown to trigger endocytosis in a wide range of cell types, including onion and potato cells ([https://academic.oup.com/pcp/article/46/3/482/1817347 Chang <i>et al.</i>]).
-If CPP can be used as a protein tag for import into an endosymbiotic symbiont, the host proteins targeted to the symbiont would simply need the CPP added.