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To identify genes driving encephalitogenic CD4+ T cell migration into the CNS, we performed a genome-wide CRISPR screen and a subsequent validation screen. For the genome-wide CRISPR screen, up to 4 sgRNA per gene and 800 non targeted controls were included, for a total of 87690 individual sgRNAs, and for the validation screen, up to 6 sgRNA per gene and 241 non targeted controls for a total of 12000 individual sgRNAs. Comparison of sgRNA distribution by pairwise comparisons across blood, spleen, ...

RNA-seq of sorted cells from brain of Cortical MS mouse treated with vehicle or CSF1R inhibitor

Astrocytes play key roles in brain function but it is still poorly understood how these are orchestrated by pan-astrocyte transcriptions factors (TFs). Here we examined the function of the well-known pan-astrocyte TF Sox9 and the novel astrocyte TF Trps1 (Transcriptional Repressor GATA Binding 1) by Cas9-mediated in vivo deletion using Mokola-pseudotyped lentiviral delivery into the adult cerebral cortex. The consequences of deleting either Sox9 or Trps1 alone or simultaneously were explored at ...

Astrocyte-specific lentiviral constructs carrying gRNAs targeting Sox9, Trps1 and or both were injected in the cerebral cortex of adult transgenic mice expressing Cas9 protein. Transduced cells were retrived from the cortex 7 and 21 days post injection and subjected to SmartSeq2 for single cell sequencing

We performed gene expression microarray analysis in the whole telencephalon at different time points during the course of regeneration after nostril or skull brain injury

MACS-sorted astrocytes, obtained from postnatal mouse spinalc cord, were transduced in vitro with Ascl1 and Neurog2. Reprogrammed cells were analyzed by electrophysiology ar 3-to-4 weeks, and then single cells were collected and subjected to SmartSeq2 amplification

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