cre dependent aav5 vector encoding channelrhodopsin paav ef1a double floxed hchr2 h134r eyfp wpre hghpa  (Addgene inc)

Journal: Nature Communications

Article Title: Multiplexed lipid nanoparticle barcoding reveals tissue-dynamic kinetic insights and enriched cellular tropism in hepatic zones

doi: 10.1038/s41467-025-68103-7

Figure Lengend Snippet: a Endogenous Rosa-tdTom fluorescent micrographs of Ai14 mouse liver, lung, and spleen 24 h post-intravenous (IV) administration of Liver SORT LNPs carrying 0.1, 0.25, or 1.0 mg/kg Cre recombinase shows liver-specific protein production. b , c Cre qPCR performed on liver, lung, and spleen 1-h ( b ) or 24-h ( c ) after administration of Liver SORT LNPs at various doses. d , e Linear regression analysis comparing tdTom signal vs. 1-h qPCR ( d ) or 24-h qPCR ( e ) shows linear correlation that is modestly stronger at 1-h. f Endogenous Rosa-tdTom fluorescent micrographs of Ai14 mouse liver, lung, and spleen 24 h post-intravenous (IV) administration of Lung SORT LNPs carrying 0.1, 0.25, or 1.0 mg/kg Cre recombinase. g , h Cre qPCR performed on liver, lung, and spleen 1-h ( g ) or 24-h ( h ) after administration of Lung SORT LNPs at various doses. i , j Linear regression analysis comparing tdTom signal vs. 1-h qPCR ( i ) or 24-h qPCR ( j ) shows linear correlation that is modestly stronger at 1-h. k Endogenous Rosa-tdTom fluorescent micrographs of Ai14 mouse liver, lung, and spleen 24 h post-intravenous (IV) administration of Spleen SORT LNPs carrying 0.1, 0.25, or 1.0 mg/kg Cre recombinase shows spleen-specific protein production. l , m Cre qPCR performed on liver, lung, and spleen 1-h ( l ) or 24-h ( m ) after administration of Liver SORT LNPs at various doses. n , o Linear regression analysis comparing tdTom signal vs. 1-hour qPCR ( n ) or 24-h qPCR ( o ) shows linear correlation that is somewhat stronger at 1-h. qPCR data ( b – e , g – j , l – o ) are presented as the mean ± SEM of N = 3 mice per dose, per timepoint, per formulation; three wells per measurement. For tdTom quantification: mean ± SEM, N = 3 image fields. Correlation analyses ( d , e , i , j , n , o ) show Pearson’s R -squared with 95% CI calculated after confirming homoscedasticity. Scale bars ( a , f , k ): 200 µm.

Article Snippet: Barcodes and a PCR adapter were cloned into an in vitro transcription (IVT) vector encoding Cre recombinase with a DNA-encoded polyA tail using Q5 SDM kit according to the manufacturer's instructions (New England Biolabs, E0554S).

Techniques: Formulation