Extended Data Fig. 11: CellREADR targeting of neuron types in rat.
From: Programmable RNA sensing for cell monitoring and manipulation
a, Schematic of binary AAV vectors for cell type targeting in rat. In READR vector, a hSyn promoter drives expression of mCherry followed by sequences coding for sesRNA, smFlag and tTA2 effectors. Along with READR, a Reporter vector drives mNeonG expression from a TRE3g promoter in response to tTA2 from the READR vector. b, Genomic structures of the rat vGAT gene with location of a sesRNA as indicated. c, AAVs READRvGAT and TRE3g-mNeon were injected into cortical deep layer and hippocampus. Binary vectors labeled cells shown in cortex (c1). c2, Magnified view of boxed region in c1. vGAT mRNAs were labeled by in-situ hybridization (c3). Co-labeling by mNeon and vGAT mRNA (c4). Arrows showed the co-labeled cells. d, Cell labeling pattern in the hippocampus CA1 region by co-injection of AAVs READRvGAT and TRE3g-mNeon. e, Magnified view of boxed region in d. Arrows indicate co-labeled cells. f–g, Specificity of rat READRvGAT in rat cortex (f) and hippocampus (g) measured as the percent of vGAT+ cells among mNeon cells. h, Genomic structures of the rat Tle4 gene with the location of a sesRNA as indicated. i–j, AAVs READRTle4 and TRE3g-mNeon co-injected into the rat motor cortex labeled cells concentrated in deep layers (i1). Tle4+ PNs were labeled by TLE4 antibody staining (i2, 3). j, Magnified view of the boxed region in i. Arrows indicate co-labeled cells by CellREADR and TLE4 antibody. k, Specificity of rat READRTle4 in rat cortex, measured as the percent of TLE4 + cells among mNeon cells. Each bar in f, g and k is the value from one rat performed (n = 1).
