Part:BBa_K5170005
A novel PROTAC using BBa_K5170003 against 231pTau
The heterobifunctional molecule is a protein targeting chimera (PROTAC) employing BBa_K5170003 selected for 231pThreonine site in hyperphosphorylated Tau proteins as the POI targeting moiety, [S,R,S]-AHPC as the ligand to localise the PROTAC to the Von Hippen Lindau E3 ubiquitin ligase and ethylene glycol as the linker bound to both the parts. Use of an aptamer instead of antibodies or other molecules in a PROTAC is due to some unconventional properties like low immunogenicity, high specificity, ease of evolution to newer targets using different forms of SELEX, selectivity to microstructure and flexibility.
Hyperphosphorylation of Tau has been associated with several neurodegenerative diseases classified as Tauopathies which even includes Alzheimer's disease. This part may be used for degradation of hyperphosphorylated Tau which is expected to stipulate further elevation of symptoms in such disorders. This PROTAC may degrade pTau depending on the presence of phosphorylation at 231 Threonine site, an AD marker found to be significantly elevated in Alzheimer’s struck human brains compared to normal brains. Moreover, the use of aptamers (due to its easy evolution) retains the scope of extrapolating this PROTAC to other sites such as 217 Threonine, 198 Serine, 235 Serine,181 Threonine, 396 Serine etc.
C8 Alkyne dCTP can be clicked with [S,R,S]-AHPC-PEG1-Azide. After HPLC purification, it can be incorporated into the BBa_K5170003 by terminal deoxynucleotidyl transferase.
Usage and Biology
The binding region of the aptamer stays undisturbed if the modified ligand linked base is joined in the 3' terminal. The modified ligand linked base can be found on https://2024.igem.wiki/iisc-bengaluru/contribution
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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