Chunk #33 — Experimental Procedures — Gibson Assembly of Vectors — lentiGuide-dTomato and lentiGuide-mTagBFP2-Hygro

Source

Evaluating Synthetic Activation and Repression of Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and Astrocytes.

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lentiGuide-Puro (Addgene, no. 52963) was digested with Mlu1 and BsiWI. dTomato was amplified from AAV-hSyn1-GCaMP6f-P2A-NLS-dTomato (Addgene, no.51085). HygroR sequence was amplified from lenti MS2-P65-HSF1_Hygro (Addgene, no. 61426). mTagBFP2 was amplified form pBAD-mTagBFP2 (Addgene, no. 3463). The P2A self-cleaving peptide sequence was amplified using a reverse primer of HygroR and forward primer of mTagBFP2. All gRNA sequences are provided in Table S2.