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crop seq car vectors  (Addgene inc)


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    Structured Review

    Addgene inc crop seq car vectors
    Crop Seq Car Vectors, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 124 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/crop+seq+guide+puro+vector/CROPseq-Guide-Puro+(Plasmid+%2386708)/pmc12545207-236-1-9
    Average 96 stars, based on 124 article reviews
    crop seq car vectors - by Bioz Stars, 2026-09
    96/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: In vivo single-cell CRISPR uncovers distinct TNF programmes in tumour evolution.
    Article Snippet: .. CROP-mCherry vector and library cloning The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites for FACS. .. The 500 sgRNA sequences were ordered as an oligonucleotide pool from IDT, cloned in batch via Gibson assembly or via BsmbI restriction sites for sgRNAs as previously described5.

    Article Title: GiRAFR improves gRNA detection and annotation in single-cell CRISPR screens.
    Article Snippet: We designed four different gRNAs for each of 25 target genes, and 20 non-targeting guides as controls using Benchling [Biology Software, 2018], retrieved from https:// benchling.com. .. Final gRNA sequences contained homology arms (5′-end: TGGAAAGGACGAAACACCG, 3′-end: GTTTT AGAGCTAGAAATAGCAAGTTAAAATAAGGC) to allow for cloning into the CROP-seq-Guide-Puro vector (addgene plasmid #86708 from Christoph Bock). .. The gRNA library was synthesized and ordered as an oligo pool through CustomArray (GenScript). gRNA cloning and lentivirus productions.

    Article Title: GiRAFR improves gRNA detection and annotation in single-cell CRISPR screens
    Article Snippet: We designed four different gRNAs for each of 25 target genes, and 20 non-targeting guides as controls using Benchling [Biology Software, 2018], retrieved from https://benchling.com . .. Final gRNA sequences contained homology arms (5′-end: TGGAAAGGACGAAACACCG, 3′-end: GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGC) to allow for cloning into the CROP-seq-Guide-Puro vector (addgene plasmid #86708 from Christoph Bock). .. The gRNA library was synthesized and ordered as an oligo pool through CustomArray (GenScript).

    Article Title: Expressed barcodes enable clonal characterization of chemotherapeutic responses in chronic lymphocytic leukemia
    Article Snippet: Cells were routinely tested for mycoplasma per the manufacturer’s instructions (VenorGeM Mycoplasma Detection Kit; Sigma-Aldrich #MP0025). .. The CROP-seq-Guide-Puro vector (Addgene, #86708) was modified by replacing the puromycin resistance gene with BFP. .. The restriction enzymes PspLI (Thermo Fisher #FERFD0854) and MluI (Thermo Fisher #FERFD0564) were used to digest and remove the puromycin resistance marker and a gBlock ® of the BFP fluorescent marker was cloned into the digested vector using the same restriction site overhangs.

    Cloning:

    Article Title: In vivo single-cell CRISPR uncovers distinct TNF programmes in tumour evolution.
    Article Snippet: .. CROP-mCherry vector and library cloning The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites for FACS. .. The 500 sgRNA sequences were ordered as an oligonucleotide pool from IDT, cloned in batch via Gibson assembly or via BsmbI restriction sites for sgRNAs as previously described5.

    Article Title: GiRAFR improves gRNA detection and annotation in single-cell CRISPR screens.
    Article Snippet: We designed four different gRNAs for each of 25 target genes, and 20 non-targeting guides as controls using Benchling [Biology Software, 2018], retrieved from https:// benchling.com. .. Final gRNA sequences contained homology arms (5′-end: TGGAAAGGACGAAACACCG, 3′-end: GTTTT AGAGCTAGAAATAGCAAGTTAAAATAAGGC) to allow for cloning into the CROP-seq-Guide-Puro vector (addgene plasmid #86708 from Christoph Bock). .. The gRNA library was synthesized and ordered as an oligo pool through CustomArray (GenScript). gRNA cloning and lentivirus productions.

    Article Title: GiRAFR improves gRNA detection and annotation in single-cell CRISPR screens
    Article Snippet: We designed four different gRNAs for each of 25 target genes, and 20 non-targeting guides as controls using Benchling [Biology Software, 2018], retrieved from https://benchling.com . .. Final gRNA sequences contained homology arms (5′-end: TGGAAAGGACGAAACACCG, 3′-end: GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGC) to allow for cloning into the CROP-seq-Guide-Puro vector (addgene plasmid #86708 from Christoph Bock). .. The gRNA library was synthesized and ordered as an oligo pool through CustomArray (GenScript).

    Sequencing:

    Article Title: In vivo single-cell CRISPR uncovers distinct TNF programmes in tumour evolution.
    Article Snippet: .. CROP-mCherry vector and library cloning The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites for FACS. .. The 500 sgRNA sequences were ordered as an oligonucleotide pool from IDT, cloned in batch via Gibson assembly or via BsmbI restriction sites for sgRNAs as previously described5.

    Article Title: In vivo single-cell CRISPR screening for microproteins identifies a critical ribosomal component
    Article Snippet: .. The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a CRE sequence, amplified from SBE-NLS-mcherry-P2A-CreERT2 PGK-rtTA3 (Addgene #196936) and cloned in via Anza 34 Pfl23II (Invitrogen IVGN0344) and Anza 28 MluI (Invitrogen IVGN0286) restriction sites. .. The NLS-mcherry-P2A fragment was removed via mutagenesis Phusion DNA Polymerase reaction (Thermo Scientific, F530S).

    Article Title: In vivo single-cell CRISPR uncovers distinct TNF programmes in tumour evolution
    Article Snippet: .. The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites for FACS. .. The 500 sgRNA sequences were ordered as an oligonucleotide pool from IDT, cloned in batch via Gibson assembly or via BsmbI restriction sites for sgRNAs as previously described .

    Article Title: In vivo single-cell ribosome profiling reveals cell-type-specific translational programs during aging
    Article Snippet: .. The ampicillin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with an mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites. ..

    Amplification:

    Article Title: In vivo single-cell CRISPR uncovers distinct TNF programmes in tumour evolution.
    Article Snippet: .. CROP-mCherry vector and library cloning The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites for FACS. .. The 500 sgRNA sequences were ordered as an oligonucleotide pool from IDT, cloned in batch via Gibson assembly or via BsmbI restriction sites for sgRNAs as previously described5.

    Article Title: In vivo single-cell CRISPR screening for microproteins identifies a critical ribosomal component
    Article Snippet: .. The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a CRE sequence, amplified from SBE-NLS-mcherry-P2A-CreERT2 PGK-rtTA3 (Addgene #196936) and cloned in via Anza 34 Pfl23II (Invitrogen IVGN0344) and Anza 28 MluI (Invitrogen IVGN0286) restriction sites. .. The NLS-mcherry-P2A fragment was removed via mutagenesis Phusion DNA Polymerase reaction (Thermo Scientific, F530S).

    Article Title: In vivo single-cell CRISPR uncovers distinct TNF programmes in tumour evolution
    Article Snippet: .. The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites for FACS. .. The 500 sgRNA sequences were ordered as an oligonucleotide pool from IDT, cloned in batch via Gibson assembly or via BsmbI restriction sites for sgRNAs as previously described .

    Article Title: In vivo single-cell ribosome profiling reveals cell-type-specific translational programs during aging
    Article Snippet: .. The ampicillin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with an mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites. ..

    Clone Assay:

    Article Title: In vivo single-cell CRISPR uncovers distinct TNF programmes in tumour evolution.
    Article Snippet: .. CROP-mCherry vector and library cloning The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites for FACS. .. The 500 sgRNA sequences were ordered as an oligonucleotide pool from IDT, cloned in batch via Gibson assembly or via BsmbI restriction sites for sgRNAs as previously described5.

    Article Title: In vivo single-cell CRISPR screening for microproteins identifies a critical ribosomal component
    Article Snippet: .. The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a CRE sequence, amplified from SBE-NLS-mcherry-P2A-CreERT2 PGK-rtTA3 (Addgene #196936) and cloned in via Anza 34 Pfl23II (Invitrogen IVGN0344) and Anza 28 MluI (Invitrogen IVGN0286) restriction sites. .. The NLS-mcherry-P2A fragment was removed via mutagenesis Phusion DNA Polymerase reaction (Thermo Scientific, F530S).

    Article Title: In vivo single-cell CRISPR uncovers distinct TNF programmes in tumour evolution
    Article Snippet: .. The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites for FACS. .. The 500 sgRNA sequences were ordered as an oligonucleotide pool from IDT, cloned in batch via Gibson assembly or via BsmbI restriction sites for sgRNAs as previously described .

    Article Title: In vivo single-cell ribosome profiling reveals cell-type-specific translational programs during aging
    Article Snippet: .. The ampicillin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with an mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites. ..

    FACS:

    Article Title: In vivo single-cell CRISPR uncovers distinct TNF programmes in tumour evolution.
    Article Snippet: .. CROP-mCherry vector and library cloning The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites for FACS. .. The 500 sgRNA sequences were ordered as an oligonucleotide pool from IDT, cloned in batch via Gibson assembly or via BsmbI restriction sites for sgRNAs as previously described5.

    Article Title: In vivo single-cell CRISPR uncovers distinct TNF programmes in tumour evolution
    Article Snippet: .. The puromycin resistance cassette in the original CROP-seq-Guide-Puro vector (Addgene #86708) was replaced with a mCherry sequence, amplified from pAAVS1-NDi-CRISPRi (Gen1) (Addgene #73497) and cloned in via Pfl23II and MluI restriction sites for FACS. .. The 500 sgRNA sequences were ordered as an oligonucleotide pool from IDT, cloned in batch via Gibson assembly or via BsmbI restriction sites for sgRNAs as previously described .

    Modification:

    Article Title: Expressed barcodes enable clonal characterization of chemotherapeutic responses in chronic lymphocytic leukemia
    Article Snippet: Cells were routinely tested for mycoplasma per the manufacturer’s instructions (VenorGeM Mycoplasma Detection Kit; Sigma-Aldrich #MP0025). .. The CROP-seq-Guide-Puro vector (Addgene, #86708) was modified by replacing the puromycin resistance gene with BFP. .. The restriction enzymes PspLI (Thermo Fisher #FERFD0854) and MluI (Thermo Fisher #FERFD0564) were used to digest and remove the puromycin resistance marker and a gBlock ® of the BFP fluorescent marker was cloned into the digested vector using the same restriction site overhangs.



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