RNA sequencing of astrocytes purified from adult brain cortices of TwKOastro mice (Knockout of Twnk in astrocytes generated with Gfap73.12:Cre driver, Jackson stock 012886)

To investigate astrocyte responses to mitochondrial dysfuncation, we induced a knockout of Twnk (encodes mitochondrial DNA helicase Twinkle) in mouse brain astrocytes (TwKOastro mice). This leads to depletion of mitochondrial genome copy number and its expression. Transcriptomes of 5 TwKOastro astrocyte preparations were compared to 6 control preparations. Astrocytes were purified from 3-3.5 months old control and TwKOastro mice using magnetic beads coated with ACSA-2 antibody (cortices from 3-4 mice per one preparation were used). RNA sequencing was done in the BGI Genomics, China. mRNA was enriched by Oligo dT selection, and reverse transcription was done using random N6 primer. End repair and adaptor ligation were followed by PCR amplification of the library. Quantification of transcript abundance was done using kallisto software, differential expression analysis was done using sleuth (PMID:27043002; PMID:28581496). TwKOastro mice = Gfap73.12Cre+;Twnkloxp/loxp or Gfap73.12Cre+;Twnkloxp/null. Control mice = Gfap73.12Cre-;Twnkloxp/loxp or Gfap73.12Cre-;Twnkloxp/null. TwKOastro mice are characterised in PMID:29302033. Generation of Twnkloxp/loxp mice is described in PMID:26924217. LoxP sites are located in the introns 1 and 3 of Twnk, Cre expression leads to a frameshift. Gfap73.12-Cre mice = JAX:012886.