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grna sequences  (Addgene inc)


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    Structured Review

    Addgene inc grna sequences
    Grna Sequences, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/grna+sequences/CROP-sgRNA-MS2+(Plasmid+%23153457)/pm41856116-865-64-67
    Average 93 stars, based on 11 article reviews
    grna sequences - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Synthesized:

    Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications.
    Article Snippet: .. The previously designed 3 gRNA sequences were synthesized with the pU6 promoter by GenScript and cloned into the lentiviral pHIV-EGFP (Addgene, #21373) plasmid14 For lentivirus production, CRISPR-Prime Editing system encoding lentiviral pLenti-PE2-BSD plasmid DNAs (Addgene, # 161514) encoding CRISPRPE containing the blasticidin resistance gene, control pHIV-EGFP non-coding pegRNA1/2/3, and coding GFP and pHIV EGFP coding pU6-pegRNA1/2/3 and GFP were transfected with pSPAX2 and pVSV-G envelope and packaging plasmid DNAs. ..

    Article Title: The cells are all-right: Regulation of the Lefty genes by separate enhancers in mouse embryonic stem cells
    Article Snippet: .. Guide RNA plasmids were assembled with gRNA sequences using the protocol described by [ ] into (Addgene 41824) or ordered as custom synthesized plasmids containing the entire gRNA expression cassette in a minimal backbone (IDT). .. F1 mouse ESCs were transfected with 5 μg of each of the 5′ gRNA(s), 3′ gRNA(s), and pCas9_GFP (Addgene 44719) [ ] plasmids using the neon or neon nxt transfection systems (Life Technologies).

    Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications
    Article Snippet: .. The previously designed 3 gRNA sequences were synthesized with the pU6 promoter by GenScript and cloned into the lentiviral pHIV-EGFP (Addgene, #21373) plasmid For lentivirus production, CRISPR-Prime Editing system encoding lentiviral pLenti-PE2-BSD plasmid DNAs (Addgene, # 161514) encoding CRISPR-PE containing the blasticidin resistance gene, control pHIV-EGFP non-coding pegRNA1/2/3, and coding GFP and pHIV EGFP coding pU6-pegRNA1/2/3 and GFP were transfected with pSPAX2 and pVSV-G envelope and packaging plasmid DNAs. ..

    Article Title: The cells are all-right: Regulation of the Lefty genes by separate enhancers in mouse embryonic stem cells.
    Article Snippet: .. Guide RNA plasmids were assembled with gRNA sequences using the protocol described by [70] into (Addgene 41824) or ordered as custom synthesized plasmids containing the entire gRNA expression cassette in a minimal backbone (IDT). .. F1 mouse ESCs were transfected with 5 μg of each of the 50 gRNA(s), 30 gRNA(s), and pCas9_GFP (Addgene 44719) [71] plasmids using the neon or neon nxt transfection systems (Life Technologies).

    Clone Assay:

    Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications.
    Article Snippet: .. The previously designed 3 gRNA sequences were synthesized with the pU6 promoter by GenScript and cloned into the lentiviral pHIV-EGFP (Addgene, #21373) plasmid14 For lentivirus production, CRISPR-Prime Editing system encoding lentiviral pLenti-PE2-BSD plasmid DNAs (Addgene, # 161514) encoding CRISPRPE containing the blasticidin resistance gene, control pHIV-EGFP non-coding pegRNA1/2/3, and coding GFP and pHIV EGFP coding pU6-pegRNA1/2/3 and GFP were transfected with pSPAX2 and pVSV-G envelope and packaging plasmid DNAs. ..

    Article Title: Mi-2β inhibitor as an immunotherapy agent
    Article Snippet: .. Validation of Genes of the Epigenetic Factors The gRNA sequences targeting the selected 18 epigenetic factors (3 gRNAs/gene) were cloned into a LentiCRISPRv2GFP vector (Addgene, #82416) following the CHOPCHOP (chopchop.rc.fas.harvard.edu) (43). .. Briefly, HEK293FT cells in 6-well plates were transfected with 1.5 μg lentiviral plasmid, 1 μg psPAX2, and 0.5 μg pMD2.G with LipofectamineTM 3000 Transfection Reagent (ThermoFisher, #L3000001).

    Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications
    Article Snippet: .. The previously designed 3 gRNA sequences were synthesized with the pU6 promoter by GenScript and cloned into the lentiviral pHIV-EGFP (Addgene, #21373) plasmid For lentivirus production, CRISPR-Prime Editing system encoding lentiviral pLenti-PE2-BSD plasmid DNAs (Addgene, # 161514) encoding CRISPR-PE containing the blasticidin resistance gene, control pHIV-EGFP non-coding pegRNA1/2/3, and coding GFP and pHIV EGFP coding pU6-pegRNA1/2/3 and GFP were transfected with pSPAX2 and pVSV-G envelope and packaging plasmid DNAs. ..

    Article Title: Cellular Feimin enhances exercise performance by suppressing muscle thermogenesis.
    Article Snippet: Exercise can rapidly increase core body temperature, and research has indicated that elevated internal body temperature can independently contribute to fatigue during physical activity.. However, the precise mechanisms responsible for regulating thermogenesis in muscles during exercise have remained unclear.. Here, we demonstrate that cellular Feimin (cFeimin) enhances exercise performance by inhibiting muscle thermogenesis during physical activity.

    CRISPR:

    Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications.
    Article Snippet: .. The previously designed 3 gRNA sequences were synthesized with the pU6 promoter by GenScript and cloned into the lentiviral pHIV-EGFP (Addgene, #21373) plasmid14 For lentivirus production, CRISPR-Prime Editing system encoding lentiviral pLenti-PE2-BSD plasmid DNAs (Addgene, # 161514) encoding CRISPRPE containing the blasticidin resistance gene, control pHIV-EGFP non-coding pegRNA1/2/3, and coding GFP and pHIV EGFP coding pU6-pegRNA1/2/3 and GFP were transfected with pSPAX2 and pVSV-G envelope and packaging plasmid DNAs. ..

    Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications
    Article Snippet: .. The previously designed 3 gRNA sequences were synthesized with the pU6 promoter by GenScript and cloned into the lentiviral pHIV-EGFP (Addgene, #21373) plasmid For lentivirus production, CRISPR-Prime Editing system encoding lentiviral pLenti-PE2-BSD plasmid DNAs (Addgene, # 161514) encoding CRISPR-PE containing the blasticidin resistance gene, control pHIV-EGFP non-coding pegRNA1/2/3, and coding GFP and pHIV EGFP coding pU6-pegRNA1/2/3 and GFP were transfected with pSPAX2 and pVSV-G envelope and packaging plasmid DNAs. ..

    Plasmid Preparation:

    Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications.
    Article Snippet: .. The previously designed 3 gRNA sequences were synthesized with the pU6 promoter by GenScript and cloned into the lentiviral pHIV-EGFP (Addgene, #21373) plasmid14 For lentivirus production, CRISPR-Prime Editing system encoding lentiviral pLenti-PE2-BSD plasmid DNAs (Addgene, # 161514) encoding CRISPRPE containing the blasticidin resistance gene, control pHIV-EGFP non-coding pegRNA1/2/3, and coding GFP and pHIV EGFP coding pU6-pegRNA1/2/3 and GFP were transfected with pSPAX2 and pVSV-G envelope and packaging plasmid DNAs. ..

    Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications
    Article Snippet: .. The previously designed 3 gRNA sequences were synthesized with the pU6 promoter by GenScript and cloned into the lentiviral pHIV-EGFP (Addgene, #21373) plasmid For lentivirus production, CRISPR-Prime Editing system encoding lentiviral pLenti-PE2-BSD plasmid DNAs (Addgene, # 161514) encoding CRISPR-PE containing the blasticidin resistance gene, control pHIV-EGFP non-coding pegRNA1/2/3, and coding GFP and pHIV EGFP coding pU6-pegRNA1/2/3 and GFP were transfected with pSPAX2 and pVSV-G envelope and packaging plasmid DNAs. ..

    Article Title: Cellular Feimin enhances exercise performance by suppressing muscle thermogenesis.
    Article Snippet: Exercise can rapidly increase core body temperature, and research has indicated that elevated internal body temperature can independently contribute to fatigue during physical activity.. However, the precise mechanisms responsible for regulating thermogenesis in muscles during exercise have remained unclear.. Here, we demonstrate that cellular Feimin (cFeimin) enhances exercise performance by inhibiting muscle thermogenesis during physical activity.

    Control:

    Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications.
    Article Snippet: .. The previously designed 3 gRNA sequences were synthesized with the pU6 promoter by GenScript and cloned into the lentiviral pHIV-EGFP (Addgene, #21373) plasmid14 For lentivirus production, CRISPR-Prime Editing system encoding lentiviral pLenti-PE2-BSD plasmid DNAs (Addgene, # 161514) encoding CRISPRPE containing the blasticidin resistance gene, control pHIV-EGFP non-coding pegRNA1/2/3, and coding GFP and pHIV EGFP coding pU6-pegRNA1/2/3 and GFP were transfected with pSPAX2 and pVSV-G envelope and packaging plasmid DNAs. ..

    Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications
    Article Snippet: .. The previously designed 3 gRNA sequences were synthesized with the pU6 promoter by GenScript and cloned into the lentiviral pHIV-EGFP (Addgene, #21373) plasmid For lentivirus production, CRISPR-Prime Editing system encoding lentiviral pLenti-PE2-BSD plasmid DNAs (Addgene, # 161514) encoding CRISPR-PE containing the blasticidin resistance gene, control pHIV-EGFP non-coding pegRNA1/2/3, and coding GFP and pHIV EGFP coding pU6-pegRNA1/2/3 and GFP were transfected with pSPAX2 and pVSV-G envelope and packaging plasmid DNAs. ..

    Transfection:

    Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications.
    Article Snippet: .. The previously designed 3 gRNA sequences were synthesized with the pU6 promoter by GenScript and cloned into the lentiviral pHIV-EGFP (Addgene, #21373) plasmid14 For lentivirus production, CRISPR-Prime Editing system encoding lentiviral pLenti-PE2-BSD plasmid DNAs (Addgene, # 161514) encoding CRISPRPE containing the blasticidin resistance gene, control pHIV-EGFP non-coding pegRNA1/2/3, and coding GFP and pHIV EGFP coding pU6-pegRNA1/2/3 and GFP were transfected with pSPAX2 and pVSV-G envelope and packaging plasmid DNAs. ..

    Article Title: CRISPR.BOT an autonomous platform for streamlined genetic engineering and molecular biology applications
    Article Snippet: .. The previously designed 3 gRNA sequences were synthesized with the pU6 promoter by GenScript and cloned into the lentiviral pHIV-EGFP (Addgene, #21373) plasmid For lentivirus production, CRISPR-Prime Editing system encoding lentiviral pLenti-PE2-BSD plasmid DNAs (Addgene, # 161514) encoding CRISPR-PE containing the blasticidin resistance gene, control pHIV-EGFP non-coding pegRNA1/2/3, and coding GFP and pHIV EGFP coding pU6-pegRNA1/2/3 and GFP were transfected with pSPAX2 and pVSV-G envelope and packaging plasmid DNAs. ..

    Biomarker Discovery:

    Article Title: Mi-2β inhibitor as an immunotherapy agent
    Article Snippet: .. Validation of Genes of the Epigenetic Factors The gRNA sequences targeting the selected 18 epigenetic factors (3 gRNAs/gene) were cloned into a LentiCRISPRv2GFP vector (Addgene, #82416) following the CHOPCHOP (chopchop.rc.fas.harvard.edu) (43). .. Briefly, HEK293FT cells in 6-well plates were transfected with 1.5 μg lentiviral plasmid, 1 μg psPAX2, and 0.5 μg pMD2.G with LipofectamineTM 3000 Transfection Reagent (ThermoFisher, #L3000001).

    Expressing:

    Article Title: The cells are all-right: Regulation of the Lefty genes by separate enhancers in mouse embryonic stem cells
    Article Snippet: .. Guide RNA plasmids were assembled with gRNA sequences using the protocol described by [ ] into (Addgene 41824) or ordered as custom synthesized plasmids containing the entire gRNA expression cassette in a minimal backbone (IDT). .. F1 mouse ESCs were transfected with 5 μg of each of the 5′ gRNA(s), 3′ gRNA(s), and pCas9_GFP (Addgene 44719) [ ] plasmids using the neon or neon nxt transfection systems (Life Technologies).

    Article Title: Functional chromatin signatures premark future lineage-specific enhancers.
    Article Snippet: .. Single-cell 10× Genomics After doxycycline-induced activation of the iSAM CRISPRa tool for 3 days, cells were washed with PBS and treated with TrypLE Select for 2 min, washed 2 times with PBS containing 0.04%BSA and filtered through CellTrics (20μm, Sysmex) to be sorted in a Flow Cytometer, using the mCherry live marker that distinguishes the transduced cell population expressing the cassette that contains the gRNA sequences (see Addgene #153457). mCherry+ hESCs transduced with the 4 different library conditions were FACSsorted and 2 cellular mixes were prepared as it follows: 1.) ..

    Article Title: The cells are all-right: Regulation of the Lefty genes by separate enhancers in mouse embryonic stem cells.
    Article Snippet: .. Guide RNA plasmids were assembled with gRNA sequences using the protocol described by [70] into (Addgene 41824) or ordered as custom synthesized plasmids containing the entire gRNA expression cassette in a minimal backbone (IDT). .. F1 mouse ESCs were transfected with 5 μg of each of the 50 gRNA(s), 30 gRNA(s), and pCas9_GFP (Addgene 44719) [71] plasmids using the neon or neon nxt transfection systems (Life Technologies).

    Single Cell:

    Article Title: Functional chromatin signatures premark future lineage-specific enhancers.
    Article Snippet: .. Single-cell 10× Genomics After doxycycline-induced activation of the iSAM CRISPRa tool for 3 days, cells were washed with PBS and treated with TrypLE Select for 2 min, washed 2 times with PBS containing 0.04%BSA and filtered through CellTrics (20μm, Sysmex) to be sorted in a Flow Cytometer, using the mCherry live marker that distinguishes the transduced cell population expressing the cassette that contains the gRNA sequences (see Addgene #153457). mCherry+ hESCs transduced with the 4 different library conditions were FACSsorted and 2 cellular mixes were prepared as it follows: 1.) ..

    Activation Assay:

    Article Title: Functional chromatin signatures premark future lineage-specific enhancers.
    Article Snippet: .. Single-cell 10× Genomics After doxycycline-induced activation of the iSAM CRISPRa tool for 3 days, cells were washed with PBS and treated with TrypLE Select for 2 min, washed 2 times with PBS containing 0.04%BSA and filtered through CellTrics (20μm, Sysmex) to be sorted in a Flow Cytometer, using the mCherry live marker that distinguishes the transduced cell population expressing the cassette that contains the gRNA sequences (see Addgene #153457). mCherry+ hESCs transduced with the 4 different library conditions were FACSsorted and 2 cellular mixes were prepared as it follows: 1.) ..

    Flow Cytometry:

    Article Title: Functional chromatin signatures premark future lineage-specific enhancers.
    Article Snippet: .. Single-cell 10× Genomics After doxycycline-induced activation of the iSAM CRISPRa tool for 3 days, cells were washed with PBS and treated with TrypLE Select for 2 min, washed 2 times with PBS containing 0.04%BSA and filtered through CellTrics (20μm, Sysmex) to be sorted in a Flow Cytometer, using the mCherry live marker that distinguishes the transduced cell population expressing the cassette that contains the gRNA sequences (see Addgene #153457). mCherry+ hESCs transduced with the 4 different library conditions were FACSsorted and 2 cellular mixes were prepared as it follows: 1.) ..

    Marker:

    Article Title: Functional chromatin signatures premark future lineage-specific enhancers.
    Article Snippet: .. Single-cell 10× Genomics After doxycycline-induced activation of the iSAM CRISPRa tool for 3 days, cells were washed with PBS and treated with TrypLE Select for 2 min, washed 2 times with PBS containing 0.04%BSA and filtered through CellTrics (20μm, Sysmex) to be sorted in a Flow Cytometer, using the mCherry live marker that distinguishes the transduced cell population expressing the cassette that contains the gRNA sequences (see Addgene #153457). mCherry+ hESCs transduced with the 4 different library conditions were FACSsorted and 2 cellular mixes were prepared as it follows: 1.) ..

    Transduction:

    Article Title: Functional chromatin signatures premark future lineage-specific enhancers.
    Article Snippet: .. Single-cell 10× Genomics After doxycycline-induced activation of the iSAM CRISPRa tool for 3 days, cells were washed with PBS and treated with TrypLE Select for 2 min, washed 2 times with PBS containing 0.04%BSA and filtered through CellTrics (20μm, Sysmex) to be sorted in a Flow Cytometer, using the mCherry live marker that distinguishes the transduced cell population expressing the cassette that contains the gRNA sequences (see Addgene #153457). mCherry+ hESCs transduced with the 4 different library conditions were FACSsorted and 2 cellular mixes were prepared as it follows: 1.) ..



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    Image Search Results


    Results of CRISPR–Cas9 plasmids for targeted gene disruption in A. fijiensis . ( a ) The sgRNA scaffold sequence was amplified from plasmid pX330 using primers gRNA-scaffold-F and gRNA-scaffold-R. The predicted endogenous U6 promoter was amplified from the A. fijiensis genome using adaptor primers U6-1-F and U6-1-R containing 5′ overlapping sequences of the sgRNA scaffold. the 20 bp pyrG -targeting sgRNA sequence was seamlessly inserted between the U6 promoter and sgRNA scaffold using primers U6- pyrG -F and U6-1- pyrG -R. The plasmid backbone containing the Cas9 open reading frame and the AMA1 autonomous replication element was amplified from plasmid FM-6 using primers pAMA-1-F and pAMA1-R. The U6- pyrG -sgRNA expression cassette was amplified from pPu6- pyrG -sgRNA using primers U6-1-F and gRNA-scaffold-R, and subsequently inserted into the Cas9-containing backbone by homologous recombination to generate the final plasmid AFM-Δ pyrG . For clarity, DNA elements in the schematic are not drawn to scale. ( b ) Targeted disruption of the pyrG gene mediated by plasmid-based CRISPR-Cas9 editing in A. fijiensis . Diagnostic PCR analysis of genomic DNA from independent transformants in pyrG locus. The DNA molecular weight marker used was a 100–5000 bp DNA Marker III (Biosharp BL103A, Anhui, China). PCR amplification was performed using primers flanking the targeted integration region to distinguish wild-type and disrupted alleles.

    Journal: Journal of Fungi

    Article Title: A High-Efficiency CRISPR–Cas9 Ribonucleoprotein Genome Editing System in Aspergillus fijiensis Enabled by Microhomology-Mediated End Joining

    doi: 10.3390/jof12030165

    Figure Lengend Snippet: Results of CRISPR–Cas9 plasmids for targeted gene disruption in A. fijiensis . ( a ) The sgRNA scaffold sequence was amplified from plasmid pX330 using primers gRNA-scaffold-F and gRNA-scaffold-R. The predicted endogenous U6 promoter was amplified from the A. fijiensis genome using adaptor primers U6-1-F and U6-1-R containing 5′ overlapping sequences of the sgRNA scaffold. the 20 bp pyrG -targeting sgRNA sequence was seamlessly inserted between the U6 promoter and sgRNA scaffold using primers U6- pyrG -F and U6-1- pyrG -R. The plasmid backbone containing the Cas9 open reading frame and the AMA1 autonomous replication element was amplified from plasmid FM-6 using primers pAMA-1-F and pAMA1-R. The U6- pyrG -sgRNA expression cassette was amplified from pPu6- pyrG -sgRNA using primers U6-1-F and gRNA-scaffold-R, and subsequently inserted into the Cas9-containing backbone by homologous recombination to generate the final plasmid AFM-Δ pyrG . For clarity, DNA elements in the schematic are not drawn to scale. ( b ) Targeted disruption of the pyrG gene mediated by plasmid-based CRISPR-Cas9 editing in A. fijiensis . Diagnostic PCR analysis of genomic DNA from independent transformants in pyrG locus. The DNA molecular weight marker used was a 100–5000 bp DNA Marker III (Biosharp BL103A, Anhui, China). PCR amplification was performed using primers flanking the targeted integration region to distinguish wild-type and disrupted alleles.

    Article Snippet: The sgRNA expression plasmid pPu6- pyrG -sgRNA was first generated by amplifying the gRNA scaffold sequence from pX330 plasmid (Addgene, Watertown, MA, USA, #42230) using primers gRNA-scaffold-F and gRNA-scaffold-R ( ).

    Techniques: CRISPR, Disruption, Sequencing, Amplification, Plasmid Preparation, Expressing, Homologous Recombination, Diagnostic Assay, Molecular Weight, Marker