biostudies-literatureGiehrl-Schwab JHelmholtz AssociationElse Kröner-Fresenius-StiftungChan Zuckerberg Initiativee14797https://www.ebi.ac.uk/biostudies/studies/S-EPMC9081909biostudies-literatureUnknown14(5)Direct reprogramming based on genetic factors resembles a promising strategy to replace lost cells in degenerative diseases such as Parkinson's disease. For this, we developed a knock-in mouse line carrying a dual dCas9 transactivator system (dCAM) allowing the conditional in vivo activation of endogenous genes. To enable a translational application, we additionally established an AAV-based strategy carrying intein-split-dCas9 in combination with activators (AAV-dCAS). Both approaches were successful in reprogramming striatal astrocytes into induced GABAergic neurons confirmed by single-cell transcriptome analysis of reprogrammed neurons in vivo. These GABAergic neurons functionally integrate into striatal circuits, alleviating voluntary motor behavior aspects in a 6-OHDA Parkinson's diseaEMBO molecular medicineParkinson's disease motor symptoms rescue by CRISPRa-reprogramming astrocytes into GABAergic neurons.PMC9081909ZT‐I‐00072018/T20xNet‐0041‐Phase2‐3Gotz MIbarra ILLuecken MDHolter SMKoupourtidou CTheis FJNinkovic JWurst WMasserdotti GTruong DJGiesert FPrakash NLao CLVogt Weisenhorn DMLefort SHembach SRauser BFischer-Sternjak JGiehrl-Schwab JfalseParkinson's disease motor symptoms rescue by CRISPRa-reprogramming astrocytes into GABAergic neurons.Direct reprogramming based on genetic factors resembles a promising strategy to replace lost cells in degenerative diseases such as Parkinson's disease. For this, we developed a knock-in mouse line carrying a dual dCas9 transactivator system (dCAM) allowing the conditional in vivo activation of endogenous genes. To enable a translational application, we additionally established an AAV-based strategy carrying intein-split-dCas9 in combination with activators (AAV-dCAS). Both approaches were successful in reprogramming striatal astrocytes into induced GABAergic neurons confirmed by single-cell transcriptome analysis of reprogrammed neurons in vivo. These GABAergic neurons functionally integrate into striatal circuits, alleviating voluntary motor behavior aspects in a 6-OHDA Parkinson's disea2022-01-01T00:00:00Z2022 May2025-04-04T20:35:41.655Z2025-02-19T03:58:46.186ZS-EPMC90819093537346410.15252/emmm.202114797