Part:BBa_K4586019
G4S linker
Part Description
This part codes for the poly-Glycine-Serine (G4S) linker peptide that is a type of flexible,unstructured synthetic peptide linker sequence. G4S linker is composed of a core pentapeptide Gly-Gly-Gly-Gly-Ser that is repeated and usually found within scFV-based synthetic receptors as CARs and syn notch receptors.
Usage
This part implemented in our design is a linker peptide to link between the elements of our internal domain the syn notch receptor which are the zinc finger peptide (ZF21.16) and, SV40 NLS and VP64 that represent our potent transcription module.
Literature Characterization
The study used G4S Linker (E7O2V) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. The Engineered Cell Line was provided by the Lohmueller Lab, University of Pittsburgh. To express flow cytometry for testing of the G4S Linker.
The study used express flow cytometry to assess live Jurkat cells, which appear blue (negative), and engineered Jurkat cells, which appear green (positive). To express an scFv-based anti-CD19 CAR containing a G4S linker.
Flow cytometry was used to watch the cell binding of GLPB30 antibodies to CAR T-cell constructs containing G4S-linked ScFvs. (A) By using an anti-human PE secondary antibody after an anti-G4S GLPB30 antibody, they could find 3–23/B3 CAR-transgene expression. Two days after CD3+ pan-T cell mRNA transduction, staining was carried out. (B) N-terminal MYC tags could be found on anti-CD19 CAR with an (G4S)3 linker, and they could be observed on lentiviral-transfected pan-T cells. (c) In contrast to MYC-negative cells, MYC-positive CAR T cells showed higher GLPB30 staining.
References
Köseer, Ayse Sedef, et al. "Immunotargeting of cancer stem cells." Cancers 15.5 (2023): 1608
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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