Products/Services Used | Details | Operation |
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Gene Synthesis> | To make pMito-mCherry-FRB-IRES-FKBP(III)-GBPen, a bicistronic vector to co-express mCherry-MitoTrap and 3×FKBP–GBPen via an internal ribosome entry site (IRES), a custom insert was made using gene synthesis (GenScript) and inserted into pEGFP-C1 in place of GFP at AgeI and EcoRI. | Get A Quote |
Tagging a protein of interest with GFP using genome editing is a popular approach to study protein function in cell and developmental biology. To avoid re-engineering cell lines or organisms in order to introduce additional tags, functionalized nanobodies that bind GFP can be used to extend the functionality of the GFP tag. We developed functionalized nanobodies, which we termed 'dongles', that could add, for example, an FKBP tag to a GFP-tagged protein of interest, enabling knocksideways experiments in GFP knock-in cell lines. The power of knocksideways is that it allows investigators to rapidly switch the protein from an active to an inactive state. We show that dongles allow for effective knocksideways of GF... More