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. 2024 Jun 17;14(10):3827–3842. doi: 10.7150/thno.95633

Figure 6.

Figure 6

The AAV9- and Sycp1-mediated induction of the CRISPR-CasRx system achieves RNA knockdown specifically in germ cells. (A) Schematic illustration of reporter RNA knockdown by Sycp1-induced CRISPR-CasRx. Vector#3 carrying the Sycp1 promoter encodes the Cre recombinase and non-targeting gRNAs (AAV9-Sycp1-Cre-NTG); Vector#4 carrying the Sycp1 promoter encodes the Cre recombinase and tdTomato-targeting gRNAs (AAV9-Sycp1-Cre-tdTomato). The testes from a Cre-dependent CasRx knock-in mice (Rosa-CAG-LSL-CasRx-tdTomato-WPRE) were microinjected with AAV9-Sycp1-Cre-NTG or AAV9-Sycp1-Cre-tdTomato. (B) Representative stereomicroscopic fluorescent images of testes from CasRx-knock-in mice microinjected with AAV9-Sycp1-Cre-NTG or AAV9-Sycp1-Cre-tdTomato. Scale bar: 1 mm. (C) Mean fluorescence intensity (MFI) of tdTomato in testes of CasRx knock-in mice microinjected with AAV9-Sycp1-Cre-NTG or AAV9-Sycp1-Cre-tdTomato. Data were normalized to the tdTomato-non-targeting condition (n = 3). (D and E) Representative flow cytometry dot plots (D) and the corresponding quantification of the percentage of tdTomato+ cells (E) in the testes of CasRx knock-in mice microinjected with AAV9-Sycp1-Cre-NTG or AAV9-Sycp1-Cre-tdTomato. Data were normalized to the tdTomato-non-targeting condition (n = 3). All data are presented as the mean ± SEM. P-values were determined using the two-tailed Student's t-test; *P < 0.05; **P < 0.01.