Fig. 3: Model prediction and the validation of the gene editing potential with top-performing mRNA LNPs.

a The Uniform Manifold Approximation and Projection (UMAP) plot of the predicted mRNA transfection potency for lipids. b Headgroups distribution and c tail combinations distribution in HeLa. d Transfection test of 15 lipid candidates selected by AI-Guided Ionizable Lipid Engineering (AGILE) platform in HeLa cells (n = 2). e Illustration of in vivo transfection study: mRNA transfection potency (mFluc) LNPs were intramuscularly injected into the mice followed by IVIS imaging at 6 h post-injection. f IVIS imaging of injection sites and livers collected from mice after intramuscular injection of mFluc LNPs, containing H9, MC3, and ALC-0315 were injected intramuscularly into mice respectively (n = 3 biologically independent mice/group, 0.5 mg kg⁻¹ mFLuc per mouse). g Quantification of luminescent intensity at the intramuscular injection site in mice (n = 3 biologically independent mice/group). h Quantification of luminescent intensity in mouse livers (n = 3 biologically independent mice/group). i Illustration of the mTmG (membrane-Tomato/membrane-Green) mouse model: in the absence of Cre recombinase, tdTomato (tandem dimer Tomato) reporter is expressed; the presence of Cre recombinase deletes STOP cassettes and results in the expression of green fluorescent protein (GFP) reporter in mice. j Representative confocal microscopy images and quantification of tdTomato and GFP expression in histological muscle and liver sections of mTmG mice post-injection of Cre mRNA loaded LNPs by intramuscular injection. Scale bar: 50 µm. Sections from 3 mice (n = 5). Red represents tdTomato, Green represents GFP, and blue represents the nucleus (DAPI). Statistical significance was analyzed by a two-way analysis of variance (ANOVA) with t-test. Source data are provided as a Source Data file. Panels (e, i) were created with BioRender.com and released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.