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human mutant lrrk2 cdna (Addgene inc)

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Addgene inc human mutant lrrk2 cdna

Characterisation of extracellular vesicles (EVs) isolated from HEK293T‐conditioned medium. (A) Schematic representation of EV‐based platform for FARM5 loading and delivery, designed to enhance BBB penetration, cellular uptake, and therapeutic efficacy in disrupting the pathogenic interaction between mutant <t>LRRK2</t> and FADD in recipient cells. (B) Size distribution and concentration analysis of HEK293T‐EVs by NTA. (C) Western blot analysis confirming the presence of EV markers CD81 and Alix in HEK293T‐EVs, while cell‐specific markers (β‐tubulin and Calnexin) were not detected. (D) A representative transmission electron microscopy (TEM) image revealing the characteristic morphology of EVs, which displays a typical biconcave‐disk shape.

Human Mutant Lrrk2 Cdna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+mutant+lrrk2+cdna/pDEST53-LRRK2-G2019S+(Plasmid+%2325045)/pmc12860401-72-0-5
Average 91 stars, based on 9 article reviews

human mutant lrrk2 cdna - by Bioz Stars, 2026-09

91/100 stars

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1) Product Images from "Extracellular Vesicle‐Mediated Delivery of Constrained Peptides Disrupts the Pathogenic Interaction of LRRK2‐FADD in Parkinson's Disease"

Article Title: Extracellular Vesicle‐Mediated Delivery of Constrained Peptides Disrupts the Pathogenic Interaction of LRRK2‐FADD in Parkinson's Disease

Journal: Journal of Extracellular Biology

doi: 10.1002/jex2.70116

Figure Legend Snippet: Characterisation of extracellular vesicles (EVs) isolated from HEK293T‐conditioned medium. (A) Schematic representation of EV‐based platform for FARM5 loading and delivery, designed to enhance BBB penetration, cellular uptake, and therapeutic efficacy in disrupting the pathogenic interaction between mutant LRRK2 and FADD in recipient cells. (B) Size distribution and concentration analysis of HEK293T‐EVs by NTA. (C) Western blot analysis confirming the presence of EV markers CD81 and Alix in HEK293T‐EVs, while cell‐specific markers (β‐tubulin and Calnexin) were not detected. (D) A representative transmission electron microscopy (TEM) image revealing the characteristic morphology of EVs, which displays a typical biconcave‐disk shape.

Techniques Used: Isolation, Drug discovery, Mutagenesis, Concentration Assay, Western Blot, Transmission Assay, Electron Microscopy

EV‐FARM5 disrupts LRRK2‐FADD interaction. (A) HEK293T cells were transiently transfected with GFP‐tagged G2019S‐LRRK2 and V5‐tagged FADD. Thirty‐six hours post‐transfection, cells were treated with vehicle, free FARM5 or EV‐FARM5 at the indicated concentrations and incubated for an additional 8 h. After treatment, cells were washed and subjected to co‐immunoprecipitation using GFP‐Trap Magnetic Agarose. The eluates were analysed for precipitated LRRK2 (GFP) and co‐eluting FADD (V5). Both free FARM5 and EV‐FARM5 disrupted the LRRK2‐FADD interaction; however, EV‐FARM5 achieved this effect at lower doses than free FARM5. (B) Quantification of the FADD/LRRK2 Co‐immunoprecipitation (Co‐IP) ratio. Band intensities from (A) were measured using ImageJ, normalised to precipitated LRRK2 (GFP), and plotted relative to treatment dose. Each dot represents a biological replicate, which consists of three technical replicates. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. NS, not significant.

Figure Legend Snippet: EV‐FARM5 disrupts LRRK2‐FADD interaction. (A) HEK293T cells were transiently transfected with GFP‐tagged G2019S‐LRRK2 and V5‐tagged FADD. Thirty‐six hours post‐transfection, cells were treated with vehicle, free FARM5 or EV‐FARM5 at the indicated concentrations and incubated for an additional 8 h. After treatment, cells were washed and subjected to co‐immunoprecipitation using GFP‐Trap Magnetic Agarose. The eluates were analysed for precipitated LRRK2 (GFP) and co‐eluting FADD (V5). Both free FARM5 and EV‐FARM5 disrupted the LRRK2‐FADD interaction; however, EV‐FARM5 achieved this effect at lower doses than free FARM5. (B) Quantification of the FADD/LRRK2 Co‐immunoprecipitation (Co‐IP) ratio. Band intensities from (A) were measured using ImageJ, normalised to precipitated LRRK2 (GFP), and plotted relative to treatment dose. Each dot represents a biological replicate, which consists of three technical replicates. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. NS, not significant.

Techniques Used: Transfection, Incubation, Immunoprecipitation, Co-Immunoprecipitation Assay

EV‐peptide enhances cell viability and reduces apoptosis in LRRK2‐G2019S‐expressing neural cells. Neuro‐2a cells were transiently transfected with either wild‐type (WT) or mutant (G2019S) LRRK2 and subsequently treated with EV‐FARM5 or free FARM5 at the indicated concentration for 16 h. (A) Cell viability was assessed using a CCK‐8 assay. Viability fold change was normalised to WT LRRK2‐expressing cells treated with vehicle control. (B) Apoptosis activity was measured using a Caspase‐Glo 3/7 assay. Both free FARM5 and EV‐FARM5 significantly enhanced cell viability (A) and suppressed apoptosis signaling activity (B) in LRRK2‐G2019S‐ expressing cells. However, EV‐FARM5 achieved these effects at lower doses than free FARM5. (C) Western blot analysis of caspase 3 and cleaved caspase 3 expression. EV‐FARM5 treatment reduced cleaved caspase‐3 levels compared to a higher dose of free FARM5, suggesting a more pronounced suppression of apoptotic signaling. Each dot represents a biological replicate, which consists of three technical replicates. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. NS, not significant.

Figure Legend Snippet: EV‐peptide enhances cell viability and reduces apoptosis in LRRK2‐G2019S‐expressing neural cells. Neuro‐2a cells were transiently transfected with either wild‐type (WT) or mutant (G2019S) LRRK2 and subsequently treated with EV‐FARM5 or free FARM5 at the indicated concentration for 16 h. (A) Cell viability was assessed using a CCK‐8 assay. Viability fold change was normalised to WT LRRK2‐expressing cells treated with vehicle control. (B) Apoptosis activity was measured using a Caspase‐Glo 3/7 assay. Both free FARM5 and EV‐FARM5 significantly enhanced cell viability (A) and suppressed apoptosis signaling activity (B) in LRRK2‐G2019S‐ expressing cells. However, EV‐FARM5 achieved these effects at lower doses than free FARM5. (C) Western blot analysis of caspase 3 and cleaved caspase 3 expression. EV‐FARM5 treatment reduced cleaved caspase‐3 levels compared to a higher dose of free FARM5, suggesting a more pronounced suppression of apoptotic signaling. Each dot represents a biological replicate, which consists of three technical replicates. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. NS, not significant.

Techniques Used: Expressing, Transfection, Mutagenesis, Concentration Assay, CCK-8 Assay, Control, Activity Assay, Caspase-Glo Assay, Western Blot

Mutagenesis:

Article Title: Extracellular Vesicle‐Mediated Delivery of Constrained Peptides Disrupts the Pathogenic Interaction of LRRK2‐FADD in Parkinson's Disease
Article Snippet: After co‐culture, cells were resuspended in 5% trypsin or Acutase (Sigma, MO, USA) and concentrated to one million cells per 50 μL for flow cytometry. .. Human mutant LRRK2 cDNA (#25045, Addgene) with an N‐terminal GFP epitope tag and full‐length V5‐tagged wild‐type (WT) FADD was used as described (Melachroinou et al. ). .. Based on previous studies (Antoniou et al. , Melachroinou et al. ), HEK293T cells were transfected simultaneously at 4:1 with V5‐FADD and GFP‐LRRK2 via JetPEI according to the manufacturer's protocol (PolyPlus transfection, NY, USA).

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Journal: Journal of Extracellular Biology

Article Title: Extracellular Vesicle‐Mediated Delivery of Constrained Peptides Disrupts the Pathogenic Interaction of LRRK2‐FADD in Parkinson's Disease

doi: 10.1002/jex2.70116

Figure Lengend Snippet: Characterisation of extracellular vesicles (EVs) isolated from HEK293T‐conditioned medium. (A) Schematic representation of EV‐based platform for FARM5 loading and delivery, designed to enhance BBB penetration, cellular uptake, and therapeutic efficacy in disrupting the pathogenic interaction between mutant LRRK2 and FADD in recipient cells. (B) Size distribution and concentration analysis of HEK293T‐EVs by NTA. (C) Western blot analysis confirming the presence of EV markers CD81 and Alix in HEK293T‐EVs, while cell‐specific markers (β‐tubulin and Calnexin) were not detected. (D) A representative transmission electron microscopy (TEM) image revealing the characteristic morphology of EVs, which displays a typical biconcave‐disk shape.

Article Snippet: Human mutant LRRK2 cDNA (#25045, Addgene) with an N‐terminal GFP epitope tag and full‐length V5‐tagged wild‐type (WT) FADD was used as described (Melachroinou et al. ).

Techniques: Isolation, Drug discovery, Mutagenesis, Concentration Assay, Western Blot, Transmission Assay, Electron Microscopy

Journal: Journal of Extracellular Biology

Article Title: Extracellular Vesicle‐Mediated Delivery of Constrained Peptides Disrupts the Pathogenic Interaction of LRRK2‐FADD in Parkinson's Disease

doi: 10.1002/jex2.70116

Figure Lengend Snippet: EV‐FARM5 disrupts LRRK2‐FADD interaction. (A) HEK293T cells were transiently transfected with GFP‐tagged G2019S‐LRRK2 and V5‐tagged FADD. Thirty‐six hours post‐transfection, cells were treated with vehicle, free FARM5 or EV‐FARM5 at the indicated concentrations and incubated for an additional 8 h. After treatment, cells were washed and subjected to co‐immunoprecipitation using GFP‐Trap Magnetic Agarose. The eluates were analysed for precipitated LRRK2 (GFP) and co‐eluting FADD (V5). Both free FARM5 and EV‐FARM5 disrupted the LRRK2‐FADD interaction; however, EV‐FARM5 achieved this effect at lower doses than free FARM5. (B) Quantification of the FADD/LRRK2 Co‐immunoprecipitation (Co‐IP) ratio. Band intensities from (A) were measured using ImageJ, normalised to precipitated LRRK2 (GFP), and plotted relative to treatment dose. Each dot represents a biological replicate, which consists of three technical replicates. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. NS, not significant.

Article Snippet: Human mutant LRRK2 cDNA (#25045, Addgene) with an N‐terminal GFP epitope tag and full‐length V5‐tagged wild‐type (WT) FADD was used as described (Melachroinou et al. ).

Techniques: Transfection, Incubation, Immunoprecipitation, Co-Immunoprecipitation Assay

Journal: Journal of Extracellular Biology

Article Title: Extracellular Vesicle‐Mediated Delivery of Constrained Peptides Disrupts the Pathogenic Interaction of LRRK2‐FADD in Parkinson's Disease

doi: 10.1002/jex2.70116

Figure Lengend Snippet: EV‐peptide enhances cell viability and reduces apoptosis in LRRK2‐G2019S‐expressing neural cells. Neuro‐2a cells were transiently transfected with either wild‐type (WT) or mutant (G2019S) LRRK2 and subsequently treated with EV‐FARM5 or free FARM5 at the indicated concentration for 16 h. (A) Cell viability was assessed using a CCK‐8 assay. Viability fold change was normalised to WT LRRK2‐expressing cells treated with vehicle control. (B) Apoptosis activity was measured using a Caspase‐Glo 3/7 assay. Both free FARM5 and EV‐FARM5 significantly enhanced cell viability (A) and suppressed apoptosis signaling activity (B) in LRRK2‐G2019S‐ expressing cells. However, EV‐FARM5 achieved these effects at lower doses than free FARM5. (C) Western blot analysis of caspase 3 and cleaved caspase 3 expression. EV‐FARM5 treatment reduced cleaved caspase‐3 levels compared to a higher dose of free FARM5, suggesting a more pronounced suppression of apoptotic signaling. Each dot represents a biological replicate, which consists of three technical replicates. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. NS, not significant.

Article Snippet: Human mutant LRRK2 cDNA (#25045, Addgene) with an N‐terminal GFP epitope tag and full‐length V5‐tagged wild‐type (WT) FADD was used as described (Melachroinou et al. ).

Techniques: Expressing, Transfection, Mutagenesis, Concentration Assay, CCK-8 Assay, Control, Activity Assay, Caspase-Glo Assay, Western Blot

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human mutant lrrk2 cdna (Addgene inc)

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1) Product Images from "Extracellular Vesicle‐Mediated Delivery of Constrained Peptides Disrupts the Pathogenic Interaction of LRRK2‐FADD in Parkinson's Disease"

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