AAVs packaged with AAV-DJ capsids were used for high efficiency of in vivo neuronal infection. AAV vectors with pHelper and pRC-DJ were transfected into HEK293 cells, which were collected and lysed 72 hr later. The virus preparations were concentrated from cell lysates by fractioning with iodixanol gradient (40%) and filtering with 100,000 MWCO tube filter. The virus titer was measured by infecting HEK293 cells and adjusted to 1.0×107 infectious units/µl for brain injection. The AAV vectors used in DN-cFos experiment (Fig. 7B, S7H–S7J) express P2A-EGFP (control) or DN cFos-P2A-EGFP under the control of Synapsin-1 promoter. The AAV vectors used in CRIPSRi experiments (Fig. 7C, S7H–J) are described in the following section and also express EGFP as a marker for dissection of infected brain tissues for further assays.
