Fig. 1
From: Targeting of NLRP3 inflammasome with gene editing for the amelioration of inflammatory diseases
Screening of CLAN for mCas9/gRNA delivery to macrophages. a General scheme of this study. We first prepared a CLAN library of different surface charge or PEG density by adjusting the amount of BHEM-Chol or polymers (PEG5K-b-PLGA11K and PLGA11K). Next, we evaluated macrophage uptake of CLANCy5-siRNA by FACS. Screened CLANs were used to encapsulate Cas9-EGFP or Cas9 mRNA and gRNA. EGFP expression of CLANmCas9-EGFP/gNC and the GFP-knockout efficiency of CLANmCas9/gGFP were detected to determine which CLAN was better for macrophage gene editing. Finally, we encapsulated mCas9/gNLRP3 into screened CLAN to prepare CLANmCas9/gNLRP3 and used it for inflammatory disease treatment. b FACS analysis of the relative uptake quantity of each CLANCy5-siRNA in macrophages of mice injected with different CLANCy5-siRNA. c FACS analysis of EGFP expression in BMDMs transfected with each CLANmCas9-EGFP/gNC. d FACS analysis of the GFP knockout efficiency of Raw264.7-GFP cells transfected with each CLANmCas9/gGFP. e FACS analysis of EGFP expression in peritoneal macrophages of mice injected with each CLANmCas9-EGFP/gNC. The data are shown as the means ± SEM of n = 3 (b) or are representative of three independent experiments (c–e)
