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Protein post-translational modifications (PTMs) represent an untapped source of targets of huge biological interest and therapeutic potential. Their validation can only be indirect, because PTMs cannot be selectively hit by nucleic-acid based interference approaches. Here we describe the protocol for Post-translational Intracellular Silencing Antibody technology (P.I.S.A.), a method for the selection of intrabodies targeting the native acetylated version of proteins and their use for PTM-selective targeting and functional interference in living cells.
Keywords
intrabodies, post-translational modifications, two-hybrid, IACT, PISA, intracellular antibodies
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The authors declare no competing financial interests
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.docx
Table 1 Sequences of validated acetylase domains and linkers Sequences of validated acetylase domains and linkers
- supplement0.docx
Reagents, media, solutions, equipment Reagents, media, solutions, equipment
- supplement0.docx
PISA screening protocol PISA screening protocol
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Associated Publications
Post-translational selective intracellular silencing of acetylated proteins with de novo selected intrabodies
by Michele Chirichella, Simonetta Lisi, Marco Fantini, +7
Nature Methods (17 January, 2017)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Version History
CURRENT STATUS: Posted
Version 1
Posted 17 Jan, 2017
Community comments: 1
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Protein post-translational modifications (PTMs) represent an untapped source of targets of huge biological interest and therapeutic potential. Their validation can only be indirect, because PTMs cannot be selectively hit by nucleic-acid based interference approaches. Here we describe the protocol for Post-translational Intracellular Silencing Antibody technology (P.I.S.A.), a method for the selection of intrabodies targeting the native acetylated version of proteins and their use for PTM-selective targeting and functional interference in living cells.
Figure 1
Figure 2
The authors declare no competing financial interests
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.docx
Table 1 Sequences of validated acetylase domains and linkers Sequences of validated acetylase domains and linkers
- supplement0.docx
Reagents, media, solutions, equipment Reagents, media, solutions, equipment
- supplement0.docx
PISA screening protocol PISA screening protocol
Loading...
Associated Publications
Post-translational selective intracellular silencing of acetylated proteins with de novo selected intrabodies
by Michele Chirichella, Simonetta Lisi, Marco Fantini, +7
Nature Methods (17 January, 2017)