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aav construct expressing mecp2 target sgrna  (Addgene inc)


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    Addgene inc aav construct expressing mecp2 target sgrna
    Aav Construct Expressing Mecp2 Target Sgrna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/sgrna+expression+plasmid+construct/MECP2+(Plasmid+%23155723)/pm41372159-179-23-41
    Average 94 stars, based on 20 article reviews
    aav construct expressing mecp2 target sgrna - by Bioz Stars, 2026-10
    94/100 stars

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    Related Articles

    Bioprocessing:

    Article Title: Targeted long-read sequencing captures CRISPR editing and AAV integration outcomes in brain
    Article Snippet: For in vivo mouse studies, four different recombinant AAV (rAAV) vectors were generated by the Research Vector Core at the Raymond G. Perelman Center for Cellular and Molecular Therapeutics at The Children’s Hospital of Philadelphia. .. PX551 AAV shuttle plasmid expressed SpCas9 under the control of the neuronal-specific Mecp2 promoter and upstream of an SV40pA (PX551 was a gift from Feng Zhang; Addgene plasmid # 60957; http://n2t.net/addgene:60957; RRID:Addgene_60957).46 gRNA expression cassettes were moved into the G0619 AAV shuttle plasmid with eGFP gene under the control of the CMV promoter and upstream of an SV40pA signal (G0619 was from the University of Iowa Viral Vector Core). .. The non-targeting ctrl-gRNA sequence used was from the GeCKO v2 CRISPR screening library.47 All rAAV plasmid shuttles have AAV2 ITR sequences. rAAV vectors were produced by the standard calcium phosphate transfection method in HEK293 cells with the AdHelper plasmid, AAV1 Rep2/Cap1 packaging plasmid, and AAV shuttle plasmids with double CsCl purification.48 Vector titers were determined by ddPCR and were 1E13 vg/mL.

    Article Title: Editing DNA methylation in vivo
    Article Snippet: PCR amplified dCas9-TET1-P2A-eGFP or dCas9-DNMT3A-P2A-eGFP fragment was cloned into the Ai-9 construct (Addgene plasmid: 22799) with FseI restriction site to generate target vectors for mESC electroporation. .. The AAV construct expressing Pcsk9 target sgRNA was cloned by inserting an mCherry-KASH fragment into the backbone construct (Addgene plasmid: 60229), and the AAV construct expressing Mecp2 target sgRNA was cloned by replacing the EGFP with mCherry in the backbone construct (Addgene plasmid: 60231) . .. For cell culture experiments, sgRNAs were cloned into the pgRNA backbone (Addgene 84477) using an AarI cut site.

    Plasmid Preparation:

    Article Title: Targeted long-read sequencing captures CRISPR editing and AAV integration outcomes in brain
    Article Snippet: For in vivo mouse studies, four different recombinant AAV (rAAV) vectors were generated by the Research Vector Core at the Raymond G. Perelman Center for Cellular and Molecular Therapeutics at The Children’s Hospital of Philadelphia. .. PX551 AAV shuttle plasmid expressed SpCas9 under the control of the neuronal-specific Mecp2 promoter and upstream of an SV40pA (PX551 was a gift from Feng Zhang; Addgene plasmid # 60957; http://n2t.net/addgene:60957; RRID:Addgene_60957).46 gRNA expression cassettes were moved into the G0619 AAV shuttle plasmid with eGFP gene under the control of the CMV promoter and upstream of an SV40pA signal (G0619 was from the University of Iowa Viral Vector Core). .. The non-targeting ctrl-gRNA sequence used was from the GeCKO v2 CRISPR screening library.47 All rAAV plasmid shuttles have AAV2 ITR sequences. rAAV vectors were produced by the standard calcium phosphate transfection method in HEK293 cells with the AdHelper plasmid, AAV1 Rep2/Cap1 packaging plasmid, and AAV shuttle plasmids with double CsCl purification.48 Vector titers were determined by ddPCR and were 1E13 vg/mL.

    Article Title: CRISPR-Hybrid: A CRISPR-Mediated Intracellular Directed Evolution Platform for RNA Aptamers.
    Article Snippet: Selection plasmids were modified from pCD185 (Addgene #113317). .. Formammalian cell assays, dCas9was cloned from plasmid #156501 (Addgene), p65-HSF1 was cloned from plasmid #61423 (Addgene), and KRAB-MeCP2 was cloned from plasmid Nature Communications | (2025) 16:595 8 #167902 (Addgene). sgRNA-chimeras were modified from plasmid #61424 (Addgene). .. PCR was performed using Q5 High-Fidelity DNA polymerase from New England Biolabs (NEB M0491L).

    Article Title: Revealing function-altering MECP2 mutations in individuals with autism spectrum disorder using yeast and Drosophila
    Article Snippet: Missense variants were generated in MECP2-pENTRY either using the Q5 mutagenesis kit (New England Biolabs, USA, # E0554) in house or were made by GenScript (New Jersey, USA). .. For MECP2 expression clone construction, MECP2-pENTRY plasmids were subcloned in pAG426GAL-ccdB (a kind gift from Susan Lindquist; Addgene plasmid # 14155) or pGW-HA.attB ( Bischof et al . 2007 ) using the Gateway LR Clonase reaction (Thermo Fisher Scientific, Waltham, USA, catalog # 11791020). .. All transgenic UAS-MECP2 variants (on pGW-HA.attB) Drosophila strains used were verified by Sanger sequencing (UBC NAPS, Vancouver, Canada) or whole amplicon sequencing (Plasmidsaurus, CA, USA) ( ).

    Article Title: Revealing function-altering MECP2 mutations in individuals with autism spectrum disorder using yeast and Drosophila.
    Article Snippet: Missense variants were generated in MECP2-14 pENTRY either using the Q5 mutagenesis kit (New England Biolabs, USA, # E0554) in house, 15 or were made by GenScript (New Jersey, USA). .. For MECP2 expression clone construction, 16 MECP2-pENTRY plasmids were subcloned in pAG426GAL-ccdB (a kind gift from a gift from 17 Susan Lindquist; Addgene plasmid # 14155) or pGW-HA.attB (Bischof et al. 2007) using the 18 Gateway LR Clonase reaction (Thermo Fisher Scientific, Waltham, USA, catalog # 11791020). .. 19 All transgenic UAS-MECP2 variants (on pGW-HA.attB) Drosophila strains used were verified 20 by Sanger sequencing (UBC NAPS, Vancouver, Canada) or whole amplicon sequencing 21 (Plasmidsaurus, CA, USA) (Supplemental Data 1).

    Article Title: Editing DNA methylation in vivo
    Article Snippet: PCR amplified dCas9-TET1-P2A-eGFP or dCas9-DNMT3A-P2A-eGFP fragment was cloned into the Ai-9 construct (Addgene plasmid: 22799) with FseI restriction site to generate target vectors for mESC electroporation. .. The AAV construct expressing Pcsk9 target sgRNA was cloned by inserting an mCherry-KASH fragment into the backbone construct (Addgene plasmid: 60229), and the AAV construct expressing Mecp2 target sgRNA was cloned by replacing the EGFP with mCherry in the backbone construct (Addgene plasmid: 60231) . .. For cell culture experiments, sgRNAs were cloned into the pgRNA backbone (Addgene 84477) using an AarI cut site.

    Control:

    Article Title: Targeted long-read sequencing captures CRISPR editing and AAV integration outcomes in brain
    Article Snippet: For in vivo mouse studies, four different recombinant AAV (rAAV) vectors were generated by the Research Vector Core at the Raymond G. Perelman Center for Cellular and Molecular Therapeutics at The Children’s Hospital of Philadelphia. .. PX551 AAV shuttle plasmid expressed SpCas9 under the control of the neuronal-specific Mecp2 promoter and upstream of an SV40pA (PX551 was a gift from Feng Zhang; Addgene plasmid # 60957; http://n2t.net/addgene:60957; RRID:Addgene_60957).46 gRNA expression cassettes were moved into the G0619 AAV shuttle plasmid with eGFP gene under the control of the CMV promoter and upstream of an SV40pA signal (G0619 was from the University of Iowa Viral Vector Core). .. The non-targeting ctrl-gRNA sequence used was from the GeCKO v2 CRISPR screening library.47 All rAAV plasmid shuttles have AAV2 ITR sequences. rAAV vectors were produced by the standard calcium phosphate transfection method in HEK293 cells with the AdHelper plasmid, AAV1 Rep2/Cap1 packaging plasmid, and AAV shuttle plasmids with double CsCl purification.48 Vector titers were determined by ddPCR and were 1E13 vg/mL.

    Expressing:

    Article Title: Targeted long-read sequencing captures CRISPR editing and AAV integration outcomes in brain
    Article Snippet: For in vivo mouse studies, four different recombinant AAV (rAAV) vectors were generated by the Research Vector Core at the Raymond G. Perelman Center for Cellular and Molecular Therapeutics at The Children’s Hospital of Philadelphia. .. PX551 AAV shuttle plasmid expressed SpCas9 under the control of the neuronal-specific Mecp2 promoter and upstream of an SV40pA (PX551 was a gift from Feng Zhang; Addgene plasmid # 60957; http://n2t.net/addgene:60957; RRID:Addgene_60957).46 gRNA expression cassettes were moved into the G0619 AAV shuttle plasmid with eGFP gene under the control of the CMV promoter and upstream of an SV40pA signal (G0619 was from the University of Iowa Viral Vector Core). .. The non-targeting ctrl-gRNA sequence used was from the GeCKO v2 CRISPR screening library.47 All rAAV plasmid shuttles have AAV2 ITR sequences. rAAV vectors were produced by the standard calcium phosphate transfection method in HEK293 cells with the AdHelper plasmid, AAV1 Rep2/Cap1 packaging plasmid, and AAV shuttle plasmids with double CsCl purification.48 Vector titers were determined by ddPCR and were 1E13 vg/mL.

    Article Title: Revealing function-altering MECP2 mutations in individuals with autism spectrum disorder using yeast and Drosophila
    Article Snippet: Missense variants were generated in MECP2-pENTRY either using the Q5 mutagenesis kit (New England Biolabs, USA, # E0554) in house or were made by GenScript (New Jersey, USA). .. For MECP2 expression clone construction, MECP2-pENTRY plasmids were subcloned in pAG426GAL-ccdB (a kind gift from Susan Lindquist; Addgene plasmid # 14155) or pGW-HA.attB ( Bischof et al . 2007 ) using the Gateway LR Clonase reaction (Thermo Fisher Scientific, Waltham, USA, catalog # 11791020). .. All transgenic UAS-MECP2 variants (on pGW-HA.attB) Drosophila strains used were verified by Sanger sequencing (UBC NAPS, Vancouver, Canada) or whole amplicon sequencing (Plasmidsaurus, CA, USA) ( ).

    Article Title: Revealing function-altering MECP2 mutations in individuals with autism spectrum disorder using yeast and Drosophila.
    Article Snippet: Missense variants were generated in MECP2-14 pENTRY either using the Q5 mutagenesis kit (New England Biolabs, USA, # E0554) in house, 15 or were made by GenScript (New Jersey, USA). .. For MECP2 expression clone construction, 16 MECP2-pENTRY plasmids were subcloned in pAG426GAL-ccdB (a kind gift from a gift from 17 Susan Lindquist; Addgene plasmid # 14155) or pGW-HA.attB (Bischof et al. 2007) using the 18 Gateway LR Clonase reaction (Thermo Fisher Scientific, Waltham, USA, catalog # 11791020). .. 19 All transgenic UAS-MECP2 variants (on pGW-HA.attB) Drosophila strains used were verified 20 by Sanger sequencing (UBC NAPS, Vancouver, Canada) or whole amplicon sequencing 21 (Plasmidsaurus, CA, USA) (Supplemental Data 1).

    Article Title: Editing DNA methylation in vivo
    Article Snippet: PCR amplified dCas9-TET1-P2A-eGFP or dCas9-DNMT3A-P2A-eGFP fragment was cloned into the Ai-9 construct (Addgene plasmid: 22799) with FseI restriction site to generate target vectors for mESC electroporation. .. The AAV construct expressing Pcsk9 target sgRNA was cloned by inserting an mCherry-KASH fragment into the backbone construct (Addgene plasmid: 60229), and the AAV construct expressing Mecp2 target sgRNA was cloned by replacing the EGFP with mCherry in the backbone construct (Addgene plasmid: 60231) . .. For cell culture experiments, sgRNAs were cloned into the pgRNA backbone (Addgene 84477) using an AarI cut site.

    Article Title: Molecular Insights into Methyllysine Recognition by the Methyl-CpG-Binding Domain of MeCP2
    Article Snippet: .. The C-terminal 6xHis-tagged MeCP2-MBD domain bacterial expression construct pNIC-CH was obtained from addgene (a gift from Cheryl Arrowsmith, addgene catalog no. 162259). .. MeCP2-MBD domain mutants W104A, F132A, Y141A, F142A, and F155A were generated using the QuikChange Lightning site-directed mutagenesis kit (Agilent catalog no. 210519), and the resulting mutations were confirmed by DNA sequencing.

    Clone Assay:

    Article Title: CRISPR-Hybrid: A CRISPR-Mediated Intracellular Directed Evolution Platform for RNA Aptamers.
    Article Snippet: Selection plasmids were modified from pCD185 (Addgene #113317). .. Formammalian cell assays, dCas9was cloned from plasmid #156501 (Addgene), p65-HSF1 was cloned from plasmid #61423 (Addgene), and KRAB-MeCP2 was cloned from plasmid Nature Communications | (2025) 16:595 8 #167902 (Addgene). sgRNA-chimeras were modified from plasmid #61424 (Addgene). .. PCR was performed using Q5 High-Fidelity DNA polymerase from New England Biolabs (NEB M0491L).

    Article Title: Editing DNA methylation in vivo
    Article Snippet: PCR amplified dCas9-TET1-P2A-eGFP or dCas9-DNMT3A-P2A-eGFP fragment was cloned into the Ai-9 construct (Addgene plasmid: 22799) with FseI restriction site to generate target vectors for mESC electroporation. .. The AAV construct expressing Pcsk9 target sgRNA was cloned by inserting an mCherry-KASH fragment into the backbone construct (Addgene plasmid: 60229), and the AAV construct expressing Mecp2 target sgRNA was cloned by replacing the EGFP with mCherry in the backbone construct (Addgene plasmid: 60231) . .. For cell culture experiments, sgRNAs were cloned into the pgRNA backbone (Addgene 84477) using an AarI cut site.

    Article Title: microRNA-422a promotes HIV replication and innate immune evasion by targeting MECP2
    Article Snippet: .. To confirm that MECP2 is a target of miR-422a, a fragment of the MECP2 3′UTR containing the predicted target site was amplified and cloned downstream of the firefly luciferase gene in the pGL-3-control vector (Addgene, #212937). ..

    Modification:

    Article Title: CRISPR-Hybrid: A CRISPR-Mediated Intracellular Directed Evolution Platform for RNA Aptamers.
    Article Snippet: Selection plasmids were modified from pCD185 (Addgene #113317). .. Formammalian cell assays, dCas9was cloned from plasmid #156501 (Addgene), p65-HSF1 was cloned from plasmid #61423 (Addgene), and KRAB-MeCP2 was cloned from plasmid Nature Communications | (2025) 16:595 8 #167902 (Addgene). sgRNA-chimeras were modified from plasmid #61424 (Addgene). .. PCR was performed using Q5 High-Fidelity DNA polymerase from New England Biolabs (NEB M0491L).

    Construct:

    Article Title: Editing DNA methylation in vivo
    Article Snippet: PCR amplified dCas9-TET1-P2A-eGFP or dCas9-DNMT3A-P2A-eGFP fragment was cloned into the Ai-9 construct (Addgene plasmid: 22799) with FseI restriction site to generate target vectors for mESC electroporation. .. The AAV construct expressing Pcsk9 target sgRNA was cloned by inserting an mCherry-KASH fragment into the backbone construct (Addgene plasmid: 60229), and the AAV construct expressing Mecp2 target sgRNA was cloned by replacing the EGFP with mCherry in the backbone construct (Addgene plasmid: 60231) . .. For cell culture experiments, sgRNAs were cloned into the pgRNA backbone (Addgene 84477) using an AarI cut site.

    Article Title: Molecular Insights into Methyllysine Recognition by the Methyl-CpG-Binding Domain of MeCP2
    Article Snippet: .. The C-terminal 6xHis-tagged MeCP2-MBD domain bacterial expression construct pNIC-CH was obtained from addgene (a gift from Cheryl Arrowsmith, addgene catalog no. 162259). .. MeCP2-MBD domain mutants W104A, F132A, Y141A, F142A, and F155A were generated using the QuikChange Lightning site-directed mutagenesis kit (Agilent catalog no. 210519), and the resulting mutations were confirmed by DNA sequencing.

    Amplification:

    Article Title: microRNA-422a promotes HIV replication and innate immune evasion by targeting MECP2
    Article Snippet: .. To confirm that MECP2 is a target of miR-422a, a fragment of the MECP2 3′UTR containing the predicted target site was amplified and cloned downstream of the firefly luciferase gene in the pGL-3-control vector (Addgene, #212937). ..

    Luciferase:

    Article Title: microRNA-422a promotes HIV replication and innate immune evasion by targeting MECP2
    Article Snippet: .. To confirm that MECP2 is a target of miR-422a, a fragment of the MECP2 3′UTR containing the predicted target site was amplified and cloned downstream of the firefly luciferase gene in the pGL-3-control vector (Addgene, #212937). ..

    other:

    Article Title: Editing DNA methylation in vivo.
    Article Snippet: PCR amplified dCas9-TET1-P2A-eGFP or dCas9-DNMT3A-P2A-eGFP fragment was cloned into the Ai-9 construct (Addgene plasmid: 22799) with FseI restriction site to generate target vectors for mESC electroporation.



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