CRISPR-Cas9 vectors

Single guide RNAs were designed toward the putative Bdnf enhancer using http://crispr.mit.edu/. The sequences of the guide RNAs that we used throughout this study are (last 3 nucleotides are PAM):

The backbones for the BPK1520 vector used to express the guide RNA (U6-BsmBIcassette-Sp-sgRNA⁹⁶ was a gift from Keith Joung (Addgene 65777). Annealed oligos composing the different guide RNAs were cloned into the BsmBI site of U6-BsmBIcassette-Sp-sgRNA. The CRISPRi repressor dCas9-KRAB-MECP2⁷⁴ was a gift from Alejandro Chavez and George Church (Addgene 110821). The backbones for the lentiviral vector pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-puro and pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-GFP⁶⁹ were a gift from Charles Gersbach (Addgene 71236, 71237) and the same guides targeting the Bdnf enhancer were cloned into the BsmBI site. To generate pLV hU6-sgRNA hUbC-dCas9-VP64-T2a-puro, we cloned the VP64 sequence from pLV hUbC-VP64 dCas9 VP64-T2A-GFP (Addgene 59791⁷⁰) into the pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-puro vector (Addgene 71236⁶⁹) using NheI.

Bdnf and control EV overexpression vectors were a gift from Christian Rosenmund.⁹¹