Project description:Striatum was isolated from 12-month-old Msh3-/- mice crossed with the Q140 knock-in (KI) HD mice and their respective controls. Genomic analysis (ATACseq) was performed on 4 genotypes: WT, Msh3-/-, Het (Q140) KI, and Het (Q140) KI X Msh3-/-, 6 replicates per genotype.
Project description:As part of collaboration between UCLA and CHDI, UCLA is creating knockouts (KOs) of 123 genes, implicated in Huntington’s disease (HD) through various computational modeling efforts. Striatum and cortex were isolated from 12-month-old Msh2-/- (KO) or Msh2+/-mice crossed with the Rgs9+/- and Q140 knock-in (KI) HD mice and their respective controls. Transcriptomic analysis (RNASeq) was performed on 9 genotypes: WT, Msh2-/-, Rgs9+/- X Msh2-/-, Rgs9+/-, Het (Q140) KI X Msh2-/-, Het (Q140) KI X Rgs9+/- X Msh2+/-, Het (Q140) KI X Rgs9+/- X Msh2-/-, Het (Q140) KI X Rgs9+/-, and Het (Q140) KI, 6-8 replicates per genotype.
Project description:MicroRNAs are small non-coding RNA molecules that fine-tune diverse biological processes and are often found to be dysregulated in diseases, such as multiple sclerosis (MS). MS is an immune-mediated disease of the central nervous system characterized by demyelination, axonal loss and neurodegeneration. We have previously shown microRNA-150 (miR-150) levels to be elevated in cell-free cerebrospinal fluid (CSF) of MS patients compared to controls. We aimed to investigate the physiopathological function of miR-150 in vivo by generating miR-150 knock-out (KO) and knock-in (KI) mice using CRISPR/Cas9 technology. To specifically interrogate the role of miR-150 upon inflammation of the central nervous system (CNS), we induced experimental autoimmune encephalomyelitis (EAE), a mouse model for MS, in these newly generated mice. After induction of EAE, miR-150KO mice developed milder disease compared to WT littermate controls while miR-150KI mice presented with exacerbated EAE. Disease amelioration in miR-150KO was accompanied by decreased infiltration of CD4+ T cells in the central nervous system compared to WT and KI mice, as well as increased FoxP3+ regulatory T (Treg) cells in inguinal lymph nodes at disease priming stage. We demonstrated that Treg cells were fundamental for EAE amelioration in miR-150KO mice, as their partial depletion during EAE priming stage in miR-150KO FoxP3DTR mice restored disease incidence and severity to the levels observed in WT and KI mice. Transcriptomic profiling of CD4+ T cells isolated from miR-150KO mice revealed upregulation of genes associated with Treg cell function, but also reduced gene translation and autophagy, as compared to WT and particularly KI cells. The role of miR-150 in affecting the fate of CD4+ T cells was further supported by the grater tendency of miR-150KO CD4+ T cells to differentiate into Treg cells in-vitro. In conclusion, miR-150 deficiency favored considerably milder CNS inflammation by promoting differentiation of a more anti-inflammatory CD4+ T cell repertoire.
Project description:Striatum was isolated from 12-month-old Msh3+/- and Msh3-/- mice crossed with the Q140 knock-in (KI) HD mice and their respective controls. Transcriptomic analysis (snRNASeq) was performed on 6 genotypes: WT, Q140, Msh3+/-, Q140 X Msh3+/-, Msh3-/-, and Q140 X Msh3-/-, 4 replicates per genotype.
Project description:Cortex was isolated from 6-month-old Pms2+/- and Pms2-/- mice crossed with the Q140 knock-in (KI) HD mice and their respective controls. Transcriptomic analysis (RNASeq) was performed on 6 genotypes: WT, Q140, Pms2+/-, Q140 X Pms2+/-, Pms2-/-, and Q140 X Pms2-/-, 8 replicates per genotype.
Project description:As part of collaboration between UCLA and CHDI, UCLA is creating knockouts (KOs) of 123 genes, implicated in Huntington’s disease (HD) through various computational modeling efforts. Striatum and cortex were isolated from 6-month-old Msh2-/- (KO) mice crossed with the Rgs9+/- and Q140 knock-in (KI) HD mice and their respective controls. Transcriptomic analysis (RNASeq) was performed on 4 genotypes: WT, Het (Q140) KI X Rgs9+/-, Q140, and Het (Q140) KI X Rgs9+/- X Msh2-/-, 8 replicates per genotype.
Project description:Cerebral cortex and striatum were isolated from 6-month old Pms1+/- and Pms1-/- mice crossed with the Q140 knock-in (KI) mice and their respective controls. Transcriptomic analysis (RNASeq) was performed on 6 genotypes: WT, Q140, Psm1+/-, Q140 X Psm1+/-, Psm1-/-, and Q140 X Psm1-/-, 8 replicates per genotype.
Project description:Striatum was isolated from 6-month-old Ccdc82+/- and Ccdc82-/- mice crossed with the Q140 knock-in (KI) HD mice and their respective controls. Transcriptomic analysis (RNASeq) was performed on 6 genotypes: WT, Q140, Ccdc82+/-, Q140 X Ccdc82+/-, Ccdc82-/-, and Q140 X Ccdc82-/-, 7-8 replicates per genotype.
Project description:Striatum was isolated from 4-month-old Mlh1+/- and Mlh1-/- mice crossed with the Q140 knock-in (KI) HD mice and their respective controls. Transcriptomic analysis (RNASeq) was performed on 6 genotypes: WT, Q140, Mlh1+/-, Q140 X Mlh1+/-, Mlh1-/-, and Q140I X Mlh1-/-, 7-9 replicates per genotype.
Project description:Cortex was isolated from 4-month-old Mlh1+/- and Mlh1-/- mice crossed with the Q140 knock-in (KI) HD mice and their respective controls. Transcriptomic analysis (RNASeq) was performed on 6 genotypes: WT, Q140, Mlh1+/-, Q140 X Mlh1+/-, Mlh1-/-, and Q140I X Mlh1-/-, 7-9 replicates per genotype.
