Extended Data Fig. 7: CellREADR targeting of PNFezf2 and PNCtip2 types in mouse cortex with ADAR2 overexpression.
From: Programmable RNA sensing for cell monitoring and manipulation
a–b, Genomic structures of mouse Fezf2 (a) and Ctip2 (b) genes with locations of sesRNAs as indicated, respectively. c, Schematic of binary AAV vectors for targeting neuron types. In READRFezf2-tTA2, a hSyn promoter drives expression of ADAR2 followed by sequences coding for sesRNAFezf2(1), T2a, and tTA2 effector. In TRE3g-mRuby3, the TRE3g promoter drives mRuby3 in response to tTA2 from the READR virus. d, Image of coronal section from a Fezf2-CreER;LoxpSTOPLoxp-H2bGFP mouse brain, showing the distribution pattern of Fezf2+ PNs in S1 somatosensory cortex (d1). Co-injection of AAVs READRFezf2(1)-tTA2 and TRE3g-mRuby3 specifically labeled PNs in L5b and L6 (d2). Co-labeling by CellREADR AAVs and H2bGFP (d3) with magnified view in (d4). Arrows indicate co-labeled cells; arrowhead shows a neuron labeled by CellREADR AAVs but not by Fezf2-H2bGFP (d4). e, Specificity of READRFezf2(1)-tTA2. f, Coronal section of WT brain immuno-stained with a CTIP2 antibody, showing the distribution pattern of Ctip2+ PNs in S1 cortex (f1). Co-injection of AAVs READRCtip2(1) and TRE3g-mRuby3 specifically labeled PNs in L5b (f2). Co-labeling by CellREADR AAVs and CTIP2 antibody (f3) with magnified view in (f4). Arrows show the co-labeled cells; arrowhead showed a mis-labeled cells by READRCtip2(1) (f4). g, Specificity of READRCtip2(1). h, Axonal projection pattern of AAV READRCtip2(1)-tTA2 and TRE3g-eYFP infected PNs in S1 cortex. Representative images showing projections to striatum (h1), thalamus (h2), midbrain (h3), pons (h4) and medulla (h5) (arrows). i, Schematic locations of coronal sections are shown on the right panel. The bars in e and g are mean values, n = 2 mice performed.
