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Image Search Results
Journal: Stem Cells
Article Title: A de novo missense mutation in PPP2R5D alters dopamine pathways and morphology of iPSC-derived midbrain neurons
doi: 10.1093/stmcls/sxae068
Figure Lengend Snippet: 50nt PspdCas13b sgRNA with Tiled A-C Mismatch Do Not Edit the Endogenous E198K site in patient-derived neural stem cells. (A) Amplicon sequencing results of the relative read number of wild-type (GAA) and E198K (AAA) reads in E198K NSCs transfected with sgRNA and dPspCas13b--ADAR2DD(E488Q/T375G)-delta-984-1090 plasmid construct. (B) Amplicon sequencing results of E198K NSCs transfected with PPIB sgRNA and dPspCas13b--ADAR2DD(E488Q/T375G)-delta-984-1090 plasmid construct. Percent editing was determined as the number of edited transcript reads over not edited transcripts in the treated group and normalized to not-treated E198K NSCs. *significantly different from not treated E198K NSCs, t-test P < .05. (C-F) Amplicon sequencing results of percent editing of the E198K site in E198K NSCs following screen with sgRNA of increasing length and PspdCas13b-ADAR2DD constructs. Percent editing was determined as the number of edited transcript reads over not edited transcripts in treated group and normalized to not-treated E198K. * ** and **** significantly different from respective construct with no gRNA control, one-way ANOVA P < .05. (G—J) Amplicon sequencing results of percent editing of the E198K site in E198K NSCS following screen with sgRNA 190 and variable A-C mismatch position and PspdCas13b-ADAR2DD constructs. Percent editing was determined as the number of edited transcript reads over not edited transcripts in treated group and normalized to not-treated E198K. * ** and **** significantly different from respective construct with no gRNA control, one-way ANOVA P < .05.
Article Snippet: Four available Psp dCas13b-ADAR2DD constructs were screened: Psp dCas13b-ADAR2DD(E488Q) (Addgene plasmid #103849),
Techniques: Derivative Assay, Amplification, Sequencing, Transfection, Plasmid Preparation, Construct, Control
Journal: Stem Cells
Article Title: A de novo missense mutation in PPP2R5D alters dopamine pathways and morphology of iPSC-derived midbrain neurons
doi: 10.1093/stmcls/sxae068
Figure Lengend Snippet: Top DEGs are not rescued by PspdCas13b-ADAR2DD constructs and sgRNA 190. Relative expression of the 3 top DEGs between the isogenic and E198K NSC cell states in isogenic, not-treated E198K NSCs, E198K NSCs treated with Psp dCas13b-ADAR2DD(E488Q), Psp dCas13b-ADAR2DD(E488Q)-delta-984-1090, Psp dCas13b-ADAR2DD(E488Q/T375G), or Psp dCas13b-ADAR2DD(E488Q/T375G)-delta-984-1090. (A) RWDD28 relative expression determined by RT-qPCR. * Significantly different from total isogenic, one-way ANOVA P < .05. (B) ZNF717 relative expression determined by RT-qPCR. * Significantly different from total isogenic, one-way ANOVA P < .05. (C) CTSF relative expression determined by RT-qPCR. * Significantly different from total isogenic, one-way ANOVA P < .05.
Article Snippet: Four available Psp dCas13b-ADAR2DD constructs were screened: Psp dCas13b-ADAR2DD(E488Q) (Addgene plasmid #103849),
Techniques: Construct, Expressing, Quantitative RT-PCR
Journal: Stem Cells
Article Title: A de novo missense mutation in PPP2R5D alters dopamine pathways and morphology of iPSC-derived midbrain neurons
doi: 10.1093/stmcls/sxae068
Figure Lengend Snippet: Top DEGs are not rescued by PspdCas13b-ADAR2DD constructs and sgRNA 190. Relative expression of the 3 top DEGs between the isogenic and E198K NSC cell states in isogenic, not-treated E198K NSCs, E198K NSCs treated with Psp dCas13b-ADAR2DD(E488Q), Psp dCas13b-ADAR2DD(E488Q)-delta-984-1090, Psp dCas13b-ADAR2DD(E488Q/T375G), or Psp dCas13b-ADAR2DD(E488Q/T375G)-delta-984-1090. (A) RWDD28 relative expression determined by RT-qPCR. * Significantly different from total isogenic, one-way ANOVA P < .05. (B) ZNF717 relative expression determined by RT-qPCR. * Significantly different from total isogenic, one-way ANOVA P < .05. (C) CTSF relative expression determined by RT-qPCR. * Significantly different from total isogenic, one-way ANOVA P < .05.
Article Snippet: Four available Psp dCas13b-ADAR2DD constructs were screened:
Techniques: Construct, Expressing, Quantitative RT-PCR