Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Mar 22;21:15330338221085370. doi: 10.1177/15330338221085370

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2022

This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (https://us.sagepub.com/en-us/nam/open-access-at-sage).

PMC Copyright notice

Figure 1. — The design and characterization of EV tracking system. (A) Schematic of EV tracking system. This system consisted of donor A549 cells that expressed sgRNAs and Cas9 proteins and recipient cells that contained STOP-FP elements. (B) Schematic of the recuperation of FP expression in reporter cells. (C) Schematic of engineered EVs. FP Nb and its affinitive Nb were fused with exosomal membrane protein CD63 and Cas9 proteins, respectively. Cas9 proteins were captured by EVs through the bond of FP-FP Nb, selectively being sorted into EVs. (D) qRT-PCR analysis of sgRNA pair levels in EVs derived from wild-type A549 or donor A549 cells (n = 3 in each group). (E) Western blot analysis of Cas9 protein levels in EVs derived from wild-type A549 cells or donor A549 cells (n = 3 in each group). (F) RNA immunoprecipitation (IP) assay of exosomal Cas9 and sgRNAs derived from wild-type A549 cells or donor A549 cells. Cas9 proteins were tagged with Flag. Lysates of EVs were blotted using Flag antibody (input) or immunoprecipitated with Flag and blotted using Flag antibody (IP). IP with anti-IgG served as control. (G) sgRNAs analysis of immunoprecipitants following IP assay in (F). Abbreviations: EV: extracellulaer vesicle; FP: fluorescent protein; Nb: nanobody; sgRNAs: single-guide RNAs.