Cre-dependent hDlx AAV virus cloning

For cloning of Cre-dependent hDlx AAV virus, we amplified the hM3D-mCherry and mCherry coding sequences from the plasmid pAAV-hSyn-DIO-hM3D(Gq)-mCherry (a gift from Bryan Roth, Addgene # 44361)^{57 (link)} and pAAV-hSyn-DIO-mCherry (also a gift from Bryan Roth, Addgene # 50459) and cloned them into a pAAV-hDlx-Flex-GFP vector backbone (a gift from Gordon Fishell, Addgene # 83895)^{58 (link)} at AccI and NheI cloning sites. The coding sequence of adra1A was synthesized from Bio Basic Inc. and cloned into a pAAV-hDlx-Flex backbone. The virus was further packaged by Vigene Biosciences Company in AAVdj serotype. pAAV-EF1a-DIO-ChR2-mCherry plasmid (a gift from Karl Deisseroth, addgene # 20297) was purchased from addgene and packaged into AAVs from Vigene Biosciences Company in serotype9. All hDlx AAV viruses were diluted to the range of 10¹¹ to 10¹² viral genome per ml with virus dilution buffer containing 350 mM NaCl and 5% D-Sorbitol in PBS.