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. 2020 Oct 5;21(19):7362. doi: 10.3390/ijms21197362

Figure 1.

Figure 1

Packaging Cas proteins into EVs. (A) Provisional technology based on WW-Ndfip1 interaction. Cas protein with a WW tag can be expressed intracellularly together with Ndfip1. Overexpressed Ndfip1 mediates ubiquitination of Cas-WW and promotes its loading into EVs. (B) Nanoblade technology. Virus-like particles are generated by Cas protein fused with HIV Gag and co-expressed with Gag-Pro-Pol protein. Resulting EVs exhibit minor carry-over of cytosolic matter and effectively enter target cells. (C) Provisional technology based on the fusion of a constitutive EV membrane protein (e.g., CD63) with a dimerization domain and the use of a hybrid Cas protein with another dimerization domain. Upon signal (light or a chemical molecule), domains dimerize and Cas protein is recruited into EVs. Post-production, the signal is removed, and the Cas protein is released into the EV lumina. This picture was created in BioRender.