chrm1 overexpression vector Search Results


chrm1 overexpression vector  (Addgene inc)
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Addgene inc chrm1 overexpression vector
( A ) DOPA-mediated GPCR activation or inhibition as determined by the PRESTO-Tango reporter assay. Data points are shaded on the basis of relative expression determined using RNA-seq in MCs (FPKM). ( B ) Log fold enrichment of CRISPR gRNAs selected for or against. Controls for protumorigenic proteins included CDK9 and PCNA. GPER1 served as an internal GPCR tumor suppressor control. High-confidence hits are targets with at least five guides that are selected for (>5-fold) or against (<0.1-fold), and where those five guides represent at least 50% of total guides for that gene. ( C ) siRNA-mediated <t>CHRM1</t> depletion in A375 human melanoma in the presence of 25 μM l -DOPA and 6.25 μM carbidopa after 5 days of treatment. Technical, n = 8. ( D ) qPCR for CHRM1 mRNA in A375 after siRNA treatment confirming knockdown. Time point taken 24 hours after siRNA transfection. Technical, n = 3. ( E ) Effect of 25 μM l -DOPA and 6.25 μM carbidopa on proliferation of A375 cells in which CHRM1 was depleted using CRISPR-Cas9 versus control gRNA against green fluorescent protein (GFP). Cell number was determined at day 5. Technical, n = 8. ( F ) Low CHRM1 expression, determined via qPCR, correlates with lack of response to 25 μM l -DOPA and 6.25 μM carbidopa. n = 3. ( G ) CHRM1 overexpression (OE) in WM2664 and RPMI-7951 human melanoma (DOPA nonresponders) in the presence or absence of 25 μM l -DOPA and 6.25 μM carbidopa after 5 days of treatment. *** P = 0.0002, **** P < 0.0001 analyzed via two-way ANOVA. n = 5. ( H ) Western blot for CHRM1 in WM2664 and RPMI-7951 after transduction with either empty vector or CHRM1.
Chrm1 Overexpression Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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chrm1 overexpression vector - by Bioz Stars, 2026-03
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( A ) DOPA-mediated GPCR activation or inhibition as determined by the PRESTO-Tango reporter assay. Data points are shaded on the basis of relative expression determined using RNA-seq in MCs (FPKM). ( B ) Log fold enrichment of CRISPR gRNAs selected for or against. Controls for protumorigenic proteins included CDK9 and PCNA. GPER1 served as an internal GPCR tumor suppressor control. High-confidence hits are targets with at least five guides that are selected for (>5-fold) or against (<0.1-fold), and where those five guides represent at least 50% of total guides for that gene. ( C ) siRNA-mediated CHRM1 depletion in A375 human melanoma in the presence of 25 μM l -DOPA and 6.25 μM carbidopa after 5 days of treatment. Technical, n = 8. ( D ) qPCR for CHRM1 mRNA in A375 after siRNA treatment confirming knockdown. Time point taken 24 hours after siRNA transfection. Technical, n = 3. ( E ) Effect of 25 μM l -DOPA and 6.25 μM carbidopa on proliferation of A375 cells in which CHRM1 was depleted using CRISPR-Cas9 versus control gRNA against green fluorescent protein (GFP). Cell number was determined at day 5. Technical, n = 8. ( F ) Low CHRM1 expression, determined via qPCR, correlates with lack of response to 25 μM l -DOPA and 6.25 μM carbidopa. n = 3. ( G ) CHRM1 overexpression (OE) in WM2664 and RPMI-7951 human melanoma (DOPA nonresponders) in the presence or absence of 25 μM l -DOPA and 6.25 μM carbidopa after 5 days of treatment. *** P = 0.0002, **** P < 0.0001 analyzed via two-way ANOVA. n = 5. ( H ) Western blot for CHRM1 in WM2664 and RPMI-7951 after transduction with either empty vector or CHRM1.

Journal: Science Advances

Article Title: Endogenous DOPA inhibits melanoma through suppression of CHRM1 signaling

doi: 10.1126/sciadv.abn4007

Figure Lengend Snippet: ( A ) DOPA-mediated GPCR activation or inhibition as determined by the PRESTO-Tango reporter assay. Data points are shaded on the basis of relative expression determined using RNA-seq in MCs (FPKM). ( B ) Log fold enrichment of CRISPR gRNAs selected for or against. Controls for protumorigenic proteins included CDK9 and PCNA. GPER1 served as an internal GPCR tumor suppressor control. High-confidence hits are targets with at least five guides that are selected for (>5-fold) or against (<0.1-fold), and where those five guides represent at least 50% of total guides for that gene. ( C ) siRNA-mediated CHRM1 depletion in A375 human melanoma in the presence of 25 μM l -DOPA and 6.25 μM carbidopa after 5 days of treatment. Technical, n = 8. ( D ) qPCR for CHRM1 mRNA in A375 after siRNA treatment confirming knockdown. Time point taken 24 hours after siRNA transfection. Technical, n = 3. ( E ) Effect of 25 μM l -DOPA and 6.25 μM carbidopa on proliferation of A375 cells in which CHRM1 was depleted using CRISPR-Cas9 versus control gRNA against green fluorescent protein (GFP). Cell number was determined at day 5. Technical, n = 8. ( F ) Low CHRM1 expression, determined via qPCR, correlates with lack of response to 25 μM l -DOPA and 6.25 μM carbidopa. n = 3. ( G ) CHRM1 overexpression (OE) in WM2664 and RPMI-7951 human melanoma (DOPA nonresponders) in the presence or absence of 25 μM l -DOPA and 6.25 μM carbidopa after 5 days of treatment. *** P = 0.0002, **** P < 0.0001 analyzed via two-way ANOVA. n = 5. ( H ) Western blot for CHRM1 in WM2664 and RPMI-7951 after transduction with either empty vector or CHRM1.

Article Snippet: CHRM1 overexpression vector was cloned from a codon-optimized plasmid available on Addgene (plasmid no. 66248).

Techniques: Activation Assay, Inhibition, Reporter Assay, Expressing, RNA Sequencing, CRISPR, Control, Knockdown, Transfection, Over Expression, Western Blot, Transduction, Plasmid Preparation

Proposed mechanism of oncogenic CHRM1 signaling in melanoma. Red text denotes inhibitors of this pathway used in this manuscript.

Journal: Science Advances

Article Title: Endogenous DOPA inhibits melanoma through suppression of CHRM1 signaling

doi: 10.1126/sciadv.abn4007

Figure Lengend Snippet: Proposed mechanism of oncogenic CHRM1 signaling in melanoma. Red text denotes inhibitors of this pathway used in this manuscript.

Article Snippet: CHRM1 overexpression vector was cloned from a codon-optimized plasmid available on Addgene (plasmid no. 66248).

Techniques: