Figure 1 | Scientific Reports

Figure 1

From: Rosa26-LSL-dCas9-VPR: a versatile mouse model for tissue specific and simultaneous activation of multiple genes for drug discovery

Figure 1The alternative text for this image may have been generated using AI.

Generation and characterization of Cre-dependent dCas9-VPR-expressing mice. (A) Scheme of the Cre-dependent dCas9-VPR Rosa26 targeting vector. (B) Outline of the in vivo experiment. Rosa26-LSL-dCas9-VPR mice were iv injected with AAV8 containing LP1-Cre gene (AAV8-Cre) or 6 different gRNAs against Pcsk9 (AAV8-gPcsk9), named gPcsk9-1 to 6, at the amount of 1 × 1011 VG/mouse. 21 days later mice were sacrificed, and tissues were collected for analysis. (C) PCR representation showing LSL cassette recombination in liver tissues isolated from Rosa26-LSL-dCas9-VPR mice transduced with AAV8-Cre or AAV8-gPCSK9 alone. A representative agarose gel electrophoresis image is shown. Lane 1–3 contains amplicons obtained from tissue samples of three different mice treated with AAV8-gPcsk9 and lanes 4–6 from tissue samples of three different mice treated with AAV8-Cre. The expected size of PCR products, marker (M) and NTC (H2O) are indicated. (D) dCas9-VPR RNA expression in tissues dissected from AAV8-Cre or AAV8-gPCSK9 injected Rosa26-LSL-dCas9-VPR mice. dCas9-VPR expression can only be seen in AAV8-Cre treated Rosa26-LSL-dCas9-VPR mice liver samples. Mean values are shown as a relative quantification, with corresponding expression level of AAV-Cre treated control group as a reference (n = 10).

Back to article page