Figure 1.

iPSC-derived human cortical-like glutamatergic neurons mimicking CLN5 Batten disease. (A) Schematic of iPSC-derived human neuron generation. Three days of induction and fourteen days of differentiation and maturation generates pure human cortical-like glutamatergic neurons. (B) iPSCs with integrated neurogenin-2 can be efficiently differentiated into pure human cortical-like glutamatergic neurons expressing neuronal markers. (C) CRISPRi strategy to inhibit CLN5 in iPSC-derived human neurons. Three sgRNAs against CLN5 were transduced using lentiviruses into iPSC-derived human neurons containing dCas9 machinery. (D,E) All three sgRNAs showed significant inhibition of CLN5, both at the transcript (D) and protein levels (D,E). (E) First three lanes are control neurons, and following three lanes are CLN5i neurons. n ≥ 4, * p < 0.05, *** p < 0.001.