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<t>Schematic</t> of CRISPR KI vector design and key elements hU6: Human <t>U6</t> promoter, amplified from the pMJ117 plasmid (Addgene plasmid #85997; a gift from Jonathan Weissman), drives robust expression of the single guide RNA (sgRNA) in mammalian cells. gRNA Insertion Site: Site cleaved by the BbsI restriction enzyme to allow insertion of the sgRNA sequence of interest. Scaffold cr1: Codon-optimized sgRNA scaffold sequence, amplified from the pMJ114 plasmid (Addgene plasmid #85995; a gift from Jonathan Weissman), forms the structural component of the sgRNA and facilitates Cas9 binding. mU6: Mouse U6 promoter, amplified from the pMJ179 plasmid (Addgene plasmid #85996; a gift from Jonathan Weissman), drives expression of a second sgRNA cassette. gRNA (DRSR2): sgRNA sequence targeting the donor recognition sequence R2 (DRSR2): 5′-GCGATCGTAATCACCCGAGT-3′,used for homology-independent targeted integration.Scaffold cr2: Codon-optimized sgRNA scaffold sequence, amplified from the pMJ179 plasmid (Addgene plasmid #85996; a gift from Jonathan Weissman, used in conjunction with the DRSR2-targeting gRNA to support Cas9 function. DRSR2: Target site recognized by the gRNA: DRSR2, 5′-GCGATCGTAATCACCCGAGTGGG-3′ to enable targeted cleavage and integration of the KI cassette. Linker: Flexible amino acid linker (N-GGGGSGGGGSGGGGS-C) inserted between the targeted protein and oScarlet to minimize steric hindrance and preserve protein function. Sequence Between the Linker and oScarlet: Distinct sequences were inserted between the linker and oScarlet to preserve the correct reading frame in each construct. The sequences are as follows: ORF-0, 5′-CCTCGA-3’; ORF-1, 5′-CTCGA-3’; ORF-2, 5′-CGCTCGA-3’.oScarlet: Codon-optimized red fluorescent protein (RFP) used as a KI tag, It enables visualization of the tagged endogenous protein in live or fixed cells, amplified from the pAAV-Ef1a-oScarlet plasmid (Addgene plasmid #137135; gift from Karl Deisseroth). X: Sequence containing multiple stop codons in all three reading frames to ensure translational termination where appropriate.
Human U6 Promoter, supplied by Addgene inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human u6 promoter/product/Addgene inc
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human u6 promoter - by Bioz Stars, 2026-04
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human u6 promoter pcr based amplification pmj179 addgene rrid addgene 85996 backbone  (Addgene inc)
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Addgene inc human u6 promoter pcr based amplification pmj179 addgene rrid addgene 85996 backbone
<t>Schematic</t> of CRISPR KI vector design and key elements hU6: Human <t>U6</t> promoter, amplified from the pMJ117 plasmid (Addgene plasmid #85997; a gift from Jonathan Weissman), drives robust expression of the single guide RNA (sgRNA) in mammalian cells. gRNA Insertion Site: Site cleaved by the BbsI restriction enzyme to allow insertion of the sgRNA sequence of interest. Scaffold cr1: Codon-optimized sgRNA scaffold sequence, amplified from the pMJ114 plasmid (Addgene plasmid #85995; a gift from Jonathan Weissman), forms the structural component of the sgRNA and facilitates Cas9 binding. mU6: Mouse U6 promoter, amplified from the pMJ179 plasmid (Addgene plasmid #85996; a gift from Jonathan Weissman), drives expression of a second sgRNA cassette. gRNA (DRSR2): sgRNA sequence targeting the donor recognition sequence R2 (DRSR2): 5′-GCGATCGTAATCACCCGAGT-3′,used for homology-independent targeted integration.Scaffold cr2: Codon-optimized sgRNA scaffold sequence, amplified from the pMJ179 plasmid (Addgene plasmid #85996; a gift from Jonathan Weissman, used in conjunction with the DRSR2-targeting gRNA to support Cas9 function. DRSR2: Target site recognized by the gRNA: DRSR2, 5′-GCGATCGTAATCACCCGAGTGGG-3′ to enable targeted cleavage and integration of the KI cassette. Linker: Flexible amino acid linker (N-GGGGSGGGGSGGGGS-C) inserted between the targeted protein and oScarlet to minimize steric hindrance and preserve protein function. Sequence Between the Linker and oScarlet: Distinct sequences were inserted between the linker and oScarlet to preserve the correct reading frame in each construct. The sequences are as follows: ORF-0, 5′-CCTCGA-3’; ORF-1, 5′-CTCGA-3’; ORF-2, 5′-CGCTCGA-3’.oScarlet: Codon-optimized red fluorescent protein (RFP) used as a KI tag, It enables visualization of the tagged endogenous protein in live or fixed cells, amplified from the pAAV-Ef1a-oScarlet plasmid (Addgene plasmid #137135; gift from Karl Deisseroth). X: Sequence containing multiple stop codons in all three reading frames to ensure translational termination where appropriate.
Human U6 Promoter Pcr Based Amplification Pmj179 Addgene Rrid Addgene 85996 Backbone, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human u6 promoter pcr based amplification pmj179 addgene rrid addgene 85996 backbone/product/Addgene inc
Average 94 stars, based on 1 article reviews
human u6 promoter pcr based amplification pmj179 addgene rrid addgene 85996 backbone - by Bioz Stars, 2026-04
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sgrna, nicking sgrna, pegrna, and epegrna vectors driven by the human u6 promoter  (Addgene inc)
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Addgene inc sgrna, nicking sgrna, pegrna, and epegrna vectors driven by the human u6 promoter
<t>Schematic</t> of CRISPR KI vector design and key elements hU6: Human <t>U6</t> promoter, amplified from the pMJ117 plasmid (Addgene plasmid #85997; a gift from Jonathan Weissman), drives robust expression of the single guide RNA (sgRNA) in mammalian cells. gRNA Insertion Site: Site cleaved by the BbsI restriction enzyme to allow insertion of the sgRNA sequence of interest. Scaffold cr1: Codon-optimized sgRNA scaffold sequence, amplified from the pMJ114 plasmid (Addgene plasmid #85995; a gift from Jonathan Weissman), forms the structural component of the sgRNA and facilitates Cas9 binding. mU6: Mouse U6 promoter, amplified from the pMJ179 plasmid (Addgene plasmid #85996; a gift from Jonathan Weissman), drives expression of a second sgRNA cassette. gRNA (DRSR2): sgRNA sequence targeting the donor recognition sequence R2 (DRSR2): 5′-GCGATCGTAATCACCCGAGT-3′,used for homology-independent targeted integration.Scaffold cr2: Codon-optimized sgRNA scaffold sequence, amplified from the pMJ179 plasmid (Addgene plasmid #85996; a gift from Jonathan Weissman, used in conjunction with the DRSR2-targeting gRNA to support Cas9 function. DRSR2: Target site recognized by the gRNA: DRSR2, 5′-GCGATCGTAATCACCCGAGTGGG-3′ to enable targeted cleavage and integration of the KI cassette. Linker: Flexible amino acid linker (N-GGGGSGGGGSGGGGS-C) inserted between the targeted protein and oScarlet to minimize steric hindrance and preserve protein function. Sequence Between the Linker and oScarlet: Distinct sequences were inserted between the linker and oScarlet to preserve the correct reading frame in each construct. The sequences are as follows: ORF-0, 5′-CCTCGA-3’; ORF-1, 5′-CTCGA-3’; ORF-2, 5′-CGCTCGA-3’.oScarlet: Codon-optimized red fluorescent protein (RFP) used as a KI tag, It enables visualization of the tagged endogenous protein in live or fixed cells, amplified from the pAAV-Ef1a-oScarlet plasmid (Addgene plasmid #137135; gift from Karl Deisseroth). X: Sequence containing multiple stop codons in all three reading frames to ensure translational termination where appropriate.
Sgrna, Nicking Sgrna, Pegrna, And Epegrna Vectors Driven By The Human U6 Promoter, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sgrna, nicking sgrna, pegrna, and epegrna vectors driven by the human u6 promoter - by Bioz Stars, 2026-04
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human ef1α promoter  (Addgene inc)
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Addgene inc human ef1α promoter
<t>Schematic</t> of CRISPR KI vector design and key elements hU6: Human <t>U6</t> promoter, amplified from the pMJ117 plasmid (Addgene plasmid #85997; a gift from Jonathan Weissman), drives robust expression of the single guide RNA (sgRNA) in mammalian cells. gRNA Insertion Site: Site cleaved by the BbsI restriction enzyme to allow insertion of the sgRNA sequence of interest. Scaffold cr1: Codon-optimized sgRNA scaffold sequence, amplified from the pMJ114 plasmid (Addgene plasmid #85995; a gift from Jonathan Weissman), forms the structural component of the sgRNA and facilitates Cas9 binding. mU6: Mouse U6 promoter, amplified from the pMJ179 plasmid (Addgene plasmid #85996; a gift from Jonathan Weissman), drives expression of a second sgRNA cassette. gRNA (DRSR2): sgRNA sequence targeting the donor recognition sequence R2 (DRSR2): 5′-GCGATCGTAATCACCCGAGT-3′,used for homology-independent targeted integration.Scaffold cr2: Codon-optimized sgRNA scaffold sequence, amplified from the pMJ179 plasmid (Addgene plasmid #85996; a gift from Jonathan Weissman, used in conjunction with the DRSR2-targeting gRNA to support Cas9 function. DRSR2: Target site recognized by the gRNA: DRSR2, 5′-GCGATCGTAATCACCCGAGTGGG-3′ to enable targeted cleavage and integration of the KI cassette. Linker: Flexible amino acid linker (N-GGGGSGGGGSGGGGS-C) inserted between the targeted protein and oScarlet to minimize steric hindrance and preserve protein function. Sequence Between the Linker and oScarlet: Distinct sequences were inserted between the linker and oScarlet to preserve the correct reading frame in each construct. The sequences are as follows: ORF-0, 5′-CCTCGA-3’; ORF-1, 5′-CTCGA-3’; ORF-2, 5′-CGCTCGA-3’.oScarlet: Codon-optimized red fluorescent protein (RFP) used as a KI tag, It enables visualization of the tagged endogenous protein in live or fixed cells, amplified from the pAAV-Ef1a-oScarlet plasmid (Addgene plasmid #137135; gift from Karl Deisseroth). X: Sequence containing multiple stop codons in all three reading frames to ensure translational termination where appropriate.
Human Ef1α Promoter, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human ef1α promoter/product/Addgene inc
Average 94 stars, based on 1 article reviews
human ef1α promoter - by Bioz Stars, 2026-04
94/100 stars
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Schematic of CRISPR KI vector design and key elements hU6: Human U6 promoter, amplified from the pMJ117 plasmid (Addgene plasmid #85997; a gift from Jonathan Weissman), drives robust expression of the single guide RNA (sgRNA) in mammalian cells. gRNA Insertion Site: Site cleaved by the BbsI restriction enzyme to allow insertion of the sgRNA sequence of interest. Scaffold cr1: Codon-optimized sgRNA scaffold sequence, amplified from the pMJ114 plasmid (Addgene plasmid #85995; a gift from Jonathan Weissman), forms the structural component of the sgRNA and facilitates Cas9 binding. mU6: Mouse U6 promoter, amplified from the pMJ179 plasmid (Addgene plasmid #85996; a gift from Jonathan Weissman), drives expression of a second sgRNA cassette. gRNA (DRSR2): sgRNA sequence targeting the donor recognition sequence R2 (DRSR2): 5′-GCGATCGTAATCACCCGAGT-3′,used for homology-independent targeted integration.Scaffold cr2: Codon-optimized sgRNA scaffold sequence, amplified from the pMJ179 plasmid (Addgene plasmid #85996; a gift from Jonathan Weissman, used in conjunction with the DRSR2-targeting gRNA to support Cas9 function. DRSR2: Target site recognized by the gRNA: DRSR2, 5′-GCGATCGTAATCACCCGAGTGGG-3′ to enable targeted cleavage and integration of the KI cassette. Linker: Flexible amino acid linker (N-GGGGSGGGGSGGGGS-C) inserted between the targeted protein and oScarlet to minimize steric hindrance and preserve protein function. Sequence Between the Linker and oScarlet: Distinct sequences were inserted between the linker and oScarlet to preserve the correct reading frame in each construct. The sequences are as follows: ORF-0, 5′-CCTCGA-3’; ORF-1, 5′-CTCGA-3’; ORF-2, 5′-CGCTCGA-3’.oScarlet: Codon-optimized red fluorescent protein (RFP) used as a KI tag, It enables visualization of the tagged endogenous protein in live or fixed cells, amplified from the pAAV-Ef1a-oScarlet plasmid (Addgene plasmid #137135; gift from Karl Deisseroth). X: Sequence containing multiple stop codons in all three reading frames to ensure translational termination where appropriate.

Journal: STAR Protocols

Article Title: Protocol for CRISPR-based manipulation and visualization of endogenous α-synuclein in cultured mouse hippocampal neurons

doi: 10.1016/j.xpro.2025.103945

Figure Lengend Snippet: Schematic of CRISPR KI vector design and key elements hU6: Human U6 promoter, amplified from the pMJ117 plasmid (Addgene plasmid #85997; a gift from Jonathan Weissman), drives robust expression of the single guide RNA (sgRNA) in mammalian cells. gRNA Insertion Site: Site cleaved by the BbsI restriction enzyme to allow insertion of the sgRNA sequence of interest. Scaffold cr1: Codon-optimized sgRNA scaffold sequence, amplified from the pMJ114 plasmid (Addgene plasmid #85995; a gift from Jonathan Weissman), forms the structural component of the sgRNA and facilitates Cas9 binding. mU6: Mouse U6 promoter, amplified from the pMJ179 plasmid (Addgene plasmid #85996; a gift from Jonathan Weissman), drives expression of a second sgRNA cassette. gRNA (DRSR2): sgRNA sequence targeting the donor recognition sequence R2 (DRSR2): 5′-GCGATCGTAATCACCCGAGT-3′,used for homology-independent targeted integration.Scaffold cr2: Codon-optimized sgRNA scaffold sequence, amplified from the pMJ179 plasmid (Addgene plasmid #85996; a gift from Jonathan Weissman, used in conjunction with the DRSR2-targeting gRNA to support Cas9 function. DRSR2: Target site recognized by the gRNA: DRSR2, 5′-GCGATCGTAATCACCCGAGTGGG-3′ to enable targeted cleavage and integration of the KI cassette. Linker: Flexible amino acid linker (N-GGGGSGGGGSGGGGS-C) inserted between the targeted protein and oScarlet to minimize steric hindrance and preserve protein function. Sequence Between the Linker and oScarlet: Distinct sequences were inserted between the linker and oScarlet to preserve the correct reading frame in each construct. The sequences are as follows: ORF-0, 5′-CCTCGA-3’; ORF-1, 5′-CTCGA-3’; ORF-2, 5′-CGCTCGA-3’.oScarlet: Codon-optimized red fluorescent protein (RFP) used as a KI tag, It enables visualization of the tagged endogenous protein in live or fixed cells, amplified from the pAAV-Ef1a-oScarlet plasmid (Addgene plasmid #137135; gift from Karl Deisseroth). X: Sequence containing multiple stop codons in all three reading frames to ensure translational termination where appropriate.

Article Snippet: Schematic of CRISPR KI vector design and key elements hU6: Human U6 promoter, amplified from the pMJ117 plasmid (Addgene plasmid #85997; a gift from Jonathan Weissman), drives robust expression of the single guide RNA (sgRNA) in mammalian cells. gRNA Insertion Site: Site cleaved by the BbsI restriction enzyme to allow insertion of the sgRNA sequence of interest.

Techniques: CRISPR, Plasmid Preparation, Amplification, Expressing, Sequencing, Binding Assay, Construct