Review



super piggybac transposase expression vector  (Addgene inc)


Bioz Verified Symbol Addgene inc is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 96
      Buy from Supplier

    Structured Review

    Addgene inc super piggybac transposase expression vector
    Super Piggybac Transposase Expression Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 143 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/transposase+expression+vector/pCRISPRia-v2+(Plasmid+%2384832)/pmc12603150-26-20-27
    Average 96 stars, based on 143 article reviews
    super piggybac transposase expression vector - by Bioz Stars, 2026-09
    96/100 stars

    Images

    Related Articles

    Transfection:

    Article Title: DNA Modifications Enabling Proximity Biotinylation
    Article Snippet: HEK293T cells were transfected using TransIT-LT1 transfection reagent (Mirus, MIR2300) following the manufacturer recommended protocol with the following change: a total of 500 ng of plasmid was transfected for a single well of a 6-well plate. .. A mixture of plasmids were transfected together: 100 ng plasmid containing a puromycin resistance marker (addgene #85599), 100 ng transposase expression vector (addgene #85601), and 300 ng pJ2941. .. Addgene plasmids #85599 and #85601 were gifts from Karl Clark and Scott Fahrenkrug.

    Plasmid Preparation:

    Article Title: DNA Modifications Enabling Proximity Biotinylation
    Article Snippet: HEK293T cells were transfected using TransIT-LT1 transfection reagent (Mirus, MIR2300) following the manufacturer recommended protocol with the following change: a total of 500 ng of plasmid was transfected for a single well of a 6-well plate. .. A mixture of plasmids were transfected together: 100 ng plasmid containing a puromycin resistance marker (addgene #85599), 100 ng transposase expression vector (addgene #85601), and 300 ng pJ2941. .. Addgene plasmids #85599 and #85601 were gifts from Karl Clark and Scott Fahrenkrug.

    Marker:

    Article Title: DNA Modifications Enabling Proximity Biotinylation
    Article Snippet: HEK293T cells were transfected using TransIT-LT1 transfection reagent (Mirus, MIR2300) following the manufacturer recommended protocol with the following change: a total of 500 ng of plasmid was transfected for a single well of a 6-well plate. .. A mixture of plasmids were transfected together: 100 ng plasmid containing a puromycin resistance marker (addgene #85599), 100 ng transposase expression vector (addgene #85601), and 300 ng pJ2941. .. Addgene plasmids #85599 and #85601 were gifts from Karl Clark and Scott Fahrenkrug.

    Expressing:

    Article Title: DNA Modifications Enabling Proximity Biotinylation
    Article Snippet: HEK293T cells were transfected using TransIT-LT1 transfection reagent (Mirus, MIR2300) following the manufacturer recommended protocol with the following change: a total of 500 ng of plasmid was transfected for a single well of a 6-well plate. .. A mixture of plasmids were transfected together: 100 ng plasmid containing a puromycin resistance marker (addgene #85599), 100 ng transposase expression vector (addgene #85601), and 300 ng pJ2941. .. Addgene plasmids #85599 and #85601 were gifts from Karl Clark and Scott Fahrenkrug.



    Similar Products

    l1110 super piggybac transposase expression vector system biosciences  (Vector Laboratories)
    94
    Vector Laboratories l1110 super piggybac transposase expression vector system biosciences
    L1110 Super Piggybac Transposase Expression Vector System Biosciences, supplied by Vector Laboratories, 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/product/transposase+expression+vector/Unconjugated+Phaseolus+Vulgaris+Leucoagglutinin+(PHA-L)/10__1016_slash_j__isci__2025__114303-243-51-56
    Average 94 stars, based on 1 article reviews
    l1110 super piggybac transposase expression vector system biosciences - by Bioz Stars, 2026-09
    94/100 stars
    		  Buy from Supplier
    super piggybac transposase expression vector  (Addgene inc)
    96
    Addgene inc super piggybac transposase expression vector
    Super Piggybac Transposase Expression Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/transposase+expression+vector/pCRISPRia-v2+(Plasmid+%2384832)/pmc12603150-26-20-27
    Average 96 stars, based on 1 article reviews
    super piggybac transposase expression vector - by Bioz Stars, 2026-09
    96/100 stars
    		  Buy from Supplier
    hyperactive piggybac transposase expression vector  (Addgene inc)
    98
    Addgene inc hyperactive piggybac transposase expression vector
    (A) VIS-seq uses fluorescent in situ sequencing of abundant circular RNA barcodes to genotype cells expressing protein variants. (1) A variant library in the VIS-seq expression cassette is integrated into cells via <t>piggyBac</t> -ase. (2) Cells are fixed; barcodes are reverse transcribed, captured with a padlock probe and amplified; (3) cells are stained and imaged; (4) barcode is sequenced in situ ; (5) single cell phenotype-genotype pairs are determined using STARCall; and (6) features for each cell are extracted using CellProfiler. (B) Schematic of VIS-seq expression cassette. PB = piggyBac ; PD = padlock probe binding site; UCOE = universal chromatin opening element ; HBB IVS2 = hemoglobin subunit beta intervening sequence 2 , ; IRES = internal ribosome entry site ; WPRE = woodchuck hepatitis virus post-transcriptional regulatory element ; cHS4 = chicken beta globin locus control region hypersensitive site 4 . (C) Time-series plot of mEGFP positivity of human WTC11 iPS cells and derived cardiomyocytes. mEGFP-tagged WT histone H1.4 is expressed via the VIS-seq cassette (blue) or lentivirus (red). Two replicates are plotted. (D) Violin plot of the number of rolling circle amplification colonies per cell after one cycle of sequencing for VIS-seq expression cassette (blue) and lentivirus (red) cells from 1F at day 14. Silenced cells were excluded. Two replicates are shown. (E) Images of VIS-seq library of mEGFP-tagged lamin A variants in U2OS cells, stained with DAPI, phalloidin, WGA, and mitoprobe (left). First base in situ sequencing of the same field-of-view; magenta=guanine, blue=thymidine, yellow=adenine, and red=cytosine (right). Some cells express lamin A variants that mislocalize into aggregates. Scale bar indicates 20 μm. (F) Images of libraries of mEGFP-tagged PTEN in human WTC11 PTEN -KO inducible-NGN2 iPS cells and mEGFP-tagged H1.4 and RPS19 in human WTC11 iPS cells, stained with DAPI and phalloidin-CF568 (left). Imaging of mEGFP-tagged PTEN library in derived neurons, seven days after induction of NGN2, stained with DAPI and anti-MAP2 AF568 (right). Some cells express variants with localization defects (PTEN=nucleocytoplasmic, HIST1H1E=chromatin binding, RPS19=nucleolar-cytoplasmic). Scale bar indicates 20 μm. (G) Imaging after first base incorporation of VIS-seq libraries from (F). PTEN library contains two 8bp barcodes per variant in each cell, whereas other libraries have one 12bp barcode per cell. Nucleobase coloring identical to (C). Scale bar indicates 20 μm.
    Hyperactive Piggybac Transposase Expression Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/transposase+expression+vector/pMD2%2EG+(Plasmid+%2312259)/bio_rxiv__2025__07__03__663081-225-16-28
    Average 98 stars, based on 1 article reviews
    hyperactive piggybac transposase expression vector - by Bioz Stars, 2026-09
    98/100 stars
    		  Buy from Supplier
    super piggybac (spb) transposase expression vectors  (System Biosciences Inc)
    90
    System Biosciences Inc super piggybac (spb) transposase expression vectors
    (A) VIS-seq uses fluorescent in situ sequencing of abundant circular RNA barcodes to genotype cells expressing protein variants. (1) A variant library in the VIS-seq expression cassette is integrated into cells via <t>piggyBac</t> -ase. (2) Cells are fixed; barcodes are reverse transcribed, captured with a padlock probe and amplified; (3) cells are stained and imaged; (4) barcode is sequenced in situ ; (5) single cell phenotype-genotype pairs are determined using STARCall; and (6) features for each cell are extracted using CellProfiler. (B) Schematic of VIS-seq expression cassette. PB = piggyBac ; PD = padlock probe binding site; UCOE = universal chromatin opening element ; HBB IVS2 = hemoglobin subunit beta intervening sequence 2 , ; IRES = internal ribosome entry site ; WPRE = woodchuck hepatitis virus post-transcriptional regulatory element ; cHS4 = chicken beta globin locus control region hypersensitive site 4 . (C) Time-series plot of mEGFP positivity of human WTC11 iPS cells and derived cardiomyocytes. mEGFP-tagged WT histone H1.4 is expressed via the VIS-seq cassette (blue) or lentivirus (red). Two replicates are plotted. (D) Violin plot of the number of rolling circle amplification colonies per cell after one cycle of sequencing for VIS-seq expression cassette (blue) and lentivirus (red) cells from 1F at day 14. Silenced cells were excluded. Two replicates are shown. (E) Images of VIS-seq library of mEGFP-tagged lamin A variants in U2OS cells, stained with DAPI, phalloidin, WGA, and mitoprobe (left). First base in situ sequencing of the same field-of-view; magenta=guanine, blue=thymidine, yellow=adenine, and red=cytosine (right). Some cells express lamin A variants that mislocalize into aggregates. Scale bar indicates 20 μm. (F) Images of libraries of mEGFP-tagged PTEN in human WTC11 PTEN -KO inducible-NGN2 iPS cells and mEGFP-tagged H1.4 and RPS19 in human WTC11 iPS cells, stained with DAPI and phalloidin-CF568 (left). Imaging of mEGFP-tagged PTEN library in derived neurons, seven days after induction of NGN2, stained with DAPI and anti-MAP2 AF568 (right). Some cells express variants with localization defects (PTEN=nucleocytoplasmic, HIST1H1E=chromatin binding, RPS19=nucleolar-cytoplasmic). Scale bar indicates 20 μm. (G) Imaging after first base incorporation of VIS-seq libraries from (F). PTEN library contains two 8bp barcodes per variant in each cell, whereas other libraries have one 12bp barcode per cell. Nucleobase coloring identical to (C). Scale bar indicates 20 μm.
    Super Piggybac (Spb) Transposase Expression Vectors, supplied by System Biosciences Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/transposase+expression+vector/piggybac+plasmids/pm40494892-297-3-7
    Average 90 stars, based on 1 article reviews
    super piggybac (spb) transposase expression vectors - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier
    recombinant dna super piggybac transposase expression vector sbi system biosciences pb210pa 1 pspcas9 bb 2a gfp addgene 48138 px330 mouse 5  (Addgene inc)
    96
    Addgene inc recombinant dna super piggybac transposase expression vector sbi system biosciences pb210pa 1 pspcas9 bb 2a gfp addgene 48138 px330 mouse 5
    (A) VIS-seq uses fluorescent in situ sequencing of abundant circular RNA barcodes to genotype cells expressing protein variants. (1) A variant library in the VIS-seq expression cassette is integrated into cells via <t>piggyBac</t> -ase. (2) Cells are fixed; barcodes are reverse transcribed, captured with a padlock probe and amplified; (3) cells are stained and imaged; (4) barcode is sequenced in situ ; (5) single cell phenotype-genotype pairs are determined using STARCall; and (6) features for each cell are extracted using CellProfiler. (B) Schematic of VIS-seq expression cassette. PB = piggyBac ; PD = padlock probe binding site; UCOE = universal chromatin opening element ; HBB IVS2 = hemoglobin subunit beta intervening sequence 2 , ; IRES = internal ribosome entry site ; WPRE = woodchuck hepatitis virus post-transcriptional regulatory element ; cHS4 = chicken beta globin locus control region hypersensitive site 4 . (C) Time-series plot of mEGFP positivity of human WTC11 iPS cells and derived cardiomyocytes. mEGFP-tagged WT histone H1.4 is expressed via the VIS-seq cassette (blue) or lentivirus (red). Two replicates are plotted. (D) Violin plot of the number of rolling circle amplification colonies per cell after one cycle of sequencing for VIS-seq expression cassette (blue) and lentivirus (red) cells from 1F at day 14. Silenced cells were excluded. Two replicates are shown. (E) Images of VIS-seq library of mEGFP-tagged lamin A variants in U2OS cells, stained with DAPI, phalloidin, WGA, and mitoprobe (left). First base in situ sequencing of the same field-of-view; magenta=guanine, blue=thymidine, yellow=adenine, and red=cytosine (right). Some cells express lamin A variants that mislocalize into aggregates. Scale bar indicates 20 μm. (F) Images of libraries of mEGFP-tagged PTEN in human WTC11 PTEN -KO inducible-NGN2 iPS cells and mEGFP-tagged H1.4 and RPS19 in human WTC11 iPS cells, stained with DAPI and phalloidin-CF568 (left). Imaging of mEGFP-tagged PTEN library in derived neurons, seven days after induction of NGN2, stained with DAPI and anti-MAP2 AF568 (right). Some cells express variants with localization defects (PTEN=nucleocytoplasmic, HIST1H1E=chromatin binding, RPS19=nucleolar-cytoplasmic). Scale bar indicates 20 μm. (G) Imaging after first base incorporation of VIS-seq libraries from (F). PTEN library contains two 8bp barcodes per variant in each cell, whereas other libraries have one 12bp barcode per cell. Nucleobase coloring identical to (C). Scale bar indicates 20 μm.
    Recombinant Dna Super Piggybac Transposase Expression Vector Sbi System Biosciences Pb210pa 1 Pspcas9 Bb 2a Gfp Addgene 48138 Px330 Mouse 5, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/transposase+expression+vector/pSpCas9(BB)-2A-GFP+(PX458)+(Plasmid+%2348138)/pm40412390-201-61-73
    Average 96 stars, based on 1 article reviews
    recombinant dna super piggybac transposase expression vector sbi system biosciences pb210pa 1 pspcas9 bb 2a gfp addgene 48138 px330 mouse 5 - by Bioz Stars, 2026-09
    96/100 stars
    		  Buy from Supplier
    the transposase expression vector  (Celetrix LLC)
    90
    Celetrix LLC the transposase expression vector
    (A) VIS-seq uses fluorescent in situ sequencing of abundant circular RNA barcodes to genotype cells expressing protein variants. (1) A variant library in the VIS-seq expression cassette is integrated into cells via <t>piggyBac</t> -ase. (2) Cells are fixed; barcodes are reverse transcribed, captured with a padlock probe and amplified; (3) cells are stained and imaged; (4) barcode is sequenced in situ ; (5) single cell phenotype-genotype pairs are determined using STARCall; and (6) features for each cell are extracted using CellProfiler. (B) Schematic of VIS-seq expression cassette. PB = piggyBac ; PD = padlock probe binding site; UCOE = universal chromatin opening element ; HBB IVS2 = hemoglobin subunit beta intervening sequence 2 , ; IRES = internal ribosome entry site ; WPRE = woodchuck hepatitis virus post-transcriptional regulatory element ; cHS4 = chicken beta globin locus control region hypersensitive site 4 . (C) Time-series plot of mEGFP positivity of human WTC11 iPS cells and derived cardiomyocytes. mEGFP-tagged WT histone H1.4 is expressed via the VIS-seq cassette (blue) or lentivirus (red). Two replicates are plotted. (D) Violin plot of the number of rolling circle amplification colonies per cell after one cycle of sequencing for VIS-seq expression cassette (blue) and lentivirus (red) cells from 1F at day 14. Silenced cells were excluded. Two replicates are shown. (E) Images of VIS-seq library of mEGFP-tagged lamin A variants in U2OS cells, stained with DAPI, phalloidin, WGA, and mitoprobe (left). First base in situ sequencing of the same field-of-view; magenta=guanine, blue=thymidine, yellow=adenine, and red=cytosine (right). Some cells express lamin A variants that mislocalize into aggregates. Scale bar indicates 20 μm. (F) Images of libraries of mEGFP-tagged PTEN in human WTC11 PTEN -KO inducible-NGN2 iPS cells and mEGFP-tagged H1.4 and RPS19 in human WTC11 iPS cells, stained with DAPI and phalloidin-CF568 (left). Imaging of mEGFP-tagged PTEN library in derived neurons, seven days after induction of NGN2, stained with DAPI and anti-MAP2 AF568 (right). Some cells express variants with localization defects (PTEN=nucleocytoplasmic, HIST1H1E=chromatin binding, RPS19=nucleolar-cytoplasmic). Scale bar indicates 20 μm. (G) Imaging after first base incorporation of VIS-seq libraries from (F). PTEN library contains two 8bp barcodes per variant in each cell, whereas other libraries have one 12bp barcode per cell. Nucleobase coloring identical to (C). Scale bar indicates 20 μm.
    The Transposase Expression Vector, supplied by Celetrix LLC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/transposase+expression+vector/the+transposase+expression+vector/bio_rxiv__2025__01__23__634500-181-6-17
    Average 90 stars, based on 1 article reviews
    the transposase expression vector - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier
    gl-2045 expression vector comprising a gl-2045 expression cassette flanked by piggybac transposase targeting sequences  (Excellgene sa)
    90
    Excellgene sa gl-2045 expression vector comprising a gl-2045 expression cassette flanked by piggybac transposase targeting sequences
    (A) VIS-seq uses fluorescent in situ sequencing of abundant circular RNA barcodes to genotype cells expressing protein variants. (1) A variant library in the VIS-seq expression cassette is integrated into cells via <t>piggyBac</t> -ase. (2) Cells are fixed; barcodes are reverse transcribed, captured with a padlock probe and amplified; (3) cells are stained and imaged; (4) barcode is sequenced in situ ; (5) single cell phenotype-genotype pairs are determined using STARCall; and (6) features for each cell are extracted using CellProfiler. (B) Schematic of VIS-seq expression cassette. PB = piggyBac ; PD = padlock probe binding site; UCOE = universal chromatin opening element ; HBB IVS2 = hemoglobin subunit beta intervening sequence 2 , ; IRES = internal ribosome entry site ; WPRE = woodchuck hepatitis virus post-transcriptional regulatory element ; cHS4 = chicken beta globin locus control region hypersensitive site 4 . (C) Time-series plot of mEGFP positivity of human WTC11 iPS cells and derived cardiomyocytes. mEGFP-tagged WT histone H1.4 is expressed via the VIS-seq cassette (blue) or lentivirus (red). Two replicates are plotted. (D) Violin plot of the number of rolling circle amplification colonies per cell after one cycle of sequencing for VIS-seq expression cassette (blue) and lentivirus (red) cells from 1F at day 14. Silenced cells were excluded. Two replicates are shown. (E) Images of VIS-seq library of mEGFP-tagged lamin A variants in U2OS cells, stained with DAPI, phalloidin, WGA, and mitoprobe (left). First base in situ sequencing of the same field-of-view; magenta=guanine, blue=thymidine, yellow=adenine, and red=cytosine (right). Some cells express lamin A variants that mislocalize into aggregates. Scale bar indicates 20 μm. (F) Images of libraries of mEGFP-tagged PTEN in human WTC11 PTEN -KO inducible-NGN2 iPS cells and mEGFP-tagged H1.4 and RPS19 in human WTC11 iPS cells, stained with DAPI and phalloidin-CF568 (left). Imaging of mEGFP-tagged PTEN library in derived neurons, seven days after induction of NGN2, stained with DAPI and anti-MAP2 AF568 (right). Some cells express variants with localization defects (PTEN=nucleocytoplasmic, HIST1H1E=chromatin binding, RPS19=nucleolar-cytoplasmic). Scale bar indicates 20 μm. (G) Imaging after first base incorporation of VIS-seq libraries from (F). PTEN library contains two 8bp barcodes per variant in each cell, whereas other libraries have one 12bp barcode per cell. Nucleobase coloring identical to (C). Scale bar indicates 20 μm.
    Gl 2045 Expression Vector Comprising A Gl 2045 Expression Cassette Flanked By Piggybac Transposase Targeting Sequences, supplied by Excellgene sa, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/transposase+expression+vector/gl+2045+expression+vector+comprising+a+gl+2045+expression+cassette+flanked+by+piggybac+transposase+targeting+sequences/us12195499-943-29-13
    Average 90 stars, based on 1 article reviews
    gl-2045 expression vector comprising a gl-2045 expression cassette flanked by piggybac transposase targeting sequences - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    (A) VIS-seq uses fluorescent in situ sequencing of abundant circular RNA barcodes to genotype cells expressing protein variants. (1) A variant library in the VIS-seq expression cassette is integrated into cells via piggyBac -ase. (2) Cells are fixed; barcodes are reverse transcribed, captured with a padlock probe and amplified; (3) cells are stained and imaged; (4) barcode is sequenced in situ ; (5) single cell phenotype-genotype pairs are determined using STARCall; and (6) features for each cell are extracted using CellProfiler. (B) Schematic of VIS-seq expression cassette. PB = piggyBac ; PD = padlock probe binding site; UCOE = universal chromatin opening element ; HBB IVS2 = hemoglobin subunit beta intervening sequence 2 , ; IRES = internal ribosome entry site ; WPRE = woodchuck hepatitis virus post-transcriptional regulatory element ; cHS4 = chicken beta globin locus control region hypersensitive site 4 . (C) Time-series plot of mEGFP positivity of human WTC11 iPS cells and derived cardiomyocytes. mEGFP-tagged WT histone H1.4 is expressed via the VIS-seq cassette (blue) or lentivirus (red). Two replicates are plotted. (D) Violin plot of the number of rolling circle amplification colonies per cell after one cycle of sequencing for VIS-seq expression cassette (blue) and lentivirus (red) cells from 1F at day 14. Silenced cells were excluded. Two replicates are shown. (E) Images of VIS-seq library of mEGFP-tagged lamin A variants in U2OS cells, stained with DAPI, phalloidin, WGA, and mitoprobe (left). First base in situ sequencing of the same field-of-view; magenta=guanine, blue=thymidine, yellow=adenine, and red=cytosine (right). Some cells express lamin A variants that mislocalize into aggregates. Scale bar indicates 20 μm. (F) Images of libraries of mEGFP-tagged PTEN in human WTC11 PTEN -KO inducible-NGN2 iPS cells and mEGFP-tagged H1.4 and RPS19 in human WTC11 iPS cells, stained with DAPI and phalloidin-CF568 (left). Imaging of mEGFP-tagged PTEN library in derived neurons, seven days after induction of NGN2, stained with DAPI and anti-MAP2 AF568 (right). Some cells express variants with localization defects (PTEN=nucleocytoplasmic, HIST1H1E=chromatin binding, RPS19=nucleolar-cytoplasmic). Scale bar indicates 20 μm. (G) Imaging after first base incorporation of VIS-seq libraries from (F). PTEN library contains two 8bp barcodes per variant in each cell, whereas other libraries have one 12bp barcode per cell. Nucleobase coloring identical to (C). Scale bar indicates 20 μm.

    Journal: bioRxiv

    Article Title: Image-based, pooled phenotyping reveals multidimensional, disease-specific variant effects

    doi: 10.1101/2025.07.03.663081

    Figure Lengend Snippet: (A) VIS-seq uses fluorescent in situ sequencing of abundant circular RNA barcodes to genotype cells expressing protein variants. (1) A variant library in the VIS-seq expression cassette is integrated into cells via piggyBac -ase. (2) Cells are fixed; barcodes are reverse transcribed, captured with a padlock probe and amplified; (3) cells are stained and imaged; (4) barcode is sequenced in situ ; (5) single cell phenotype-genotype pairs are determined using STARCall; and (6) features for each cell are extracted using CellProfiler. (B) Schematic of VIS-seq expression cassette. PB = piggyBac ; PD = padlock probe binding site; UCOE = universal chromatin opening element ; HBB IVS2 = hemoglobin subunit beta intervening sequence 2 , ; IRES = internal ribosome entry site ; WPRE = woodchuck hepatitis virus post-transcriptional regulatory element ; cHS4 = chicken beta globin locus control region hypersensitive site 4 . (C) Time-series plot of mEGFP positivity of human WTC11 iPS cells and derived cardiomyocytes. mEGFP-tagged WT histone H1.4 is expressed via the VIS-seq cassette (blue) or lentivirus (red). Two replicates are plotted. (D) Violin plot of the number of rolling circle amplification colonies per cell after one cycle of sequencing for VIS-seq expression cassette (blue) and lentivirus (red) cells from 1F at day 14. Silenced cells were excluded. Two replicates are shown. (E) Images of VIS-seq library of mEGFP-tagged lamin A variants in U2OS cells, stained with DAPI, phalloidin, WGA, and mitoprobe (left). First base in situ sequencing of the same field-of-view; magenta=guanine, blue=thymidine, yellow=adenine, and red=cytosine (right). Some cells express lamin A variants that mislocalize into aggregates. Scale bar indicates 20 μm. (F) Images of libraries of mEGFP-tagged PTEN in human WTC11 PTEN -KO inducible-NGN2 iPS cells and mEGFP-tagged H1.4 and RPS19 in human WTC11 iPS cells, stained with DAPI and phalloidin-CF568 (left). Imaging of mEGFP-tagged PTEN library in derived neurons, seven days after induction of NGN2, stained with DAPI and anti-MAP2 AF568 (right). Some cells express variants with localization defects (PTEN=nucleocytoplasmic, HIST1H1E=chromatin binding, RPS19=nucleolar-cytoplasmic). Scale bar indicates 20 μm. (G) Imaging after first base incorporation of VIS-seq libraries from (F). PTEN library contains two 8bp barcodes per variant in each cell, whereas other libraries have one 12bp barcode per cell. Nucleobase coloring identical to (C). Scale bar indicates 20 μm.

    Article Snippet: pHSG299 (Takara Bio, 3299) pHR-UCOE-SFFV-Zim3-dCas9-P2A-EGFP (Addgene, 188899) piggyFlex (Addgene, 218234) lentiGuide-Puro (Addgene, 52963) pCAG-NLS-HA-Bxb1 (Addgene, 51271) Hyperactive piggyBac transposase expression vector (hyPBase; gift from Jay Shendure lab) pMD2.G (Addgene, 12259) psPAX2 (Addgene, 12260)

    Techniques: In Situ, Sequencing, Expressing, Variant Assay, Reverse Transcription, Amplification, Staining, Binding Assay, Virus, Control, Derivative Assay, Imaging

    (A) Fraction of cells with 1, 2, or 3 integrations, determined by 4-base in situ sequencing, after co-transfection of LMNA VIS-seq library DNA at different quantities (in nanograms) with plasmid encoding Piggybac-ase (at a 4-fold lower mass). (B) Imaging of day-7 neurons derived from clonal NGN2-inducible PTEN knockout lines stained for DAPI (blue) and NCAM1 (red). Scale bar indicates 20 μm. (C) Western blot of U2OS cells showing parental line (left) and LMNA knockout lines (right) stained for lamin A protein. (D) Western blot of NGN2-inducible iPS cells showing parental line (left) and PTEN knockout lines (right) stained for PTEN and phospho-AKT protein. Clonal lines C2 and C18 were used for PTEN VIS-seq experiments.

    Journal: bioRxiv

    Article Title: Image-based, pooled phenotyping reveals multidimensional, disease-specific variant effects

    doi: 10.1101/2025.07.03.663081

    Figure Lengend Snippet: (A) Fraction of cells with 1, 2, or 3 integrations, determined by 4-base in situ sequencing, after co-transfection of LMNA VIS-seq library DNA at different quantities (in nanograms) with plasmid encoding Piggybac-ase (at a 4-fold lower mass). (B) Imaging of day-7 neurons derived from clonal NGN2-inducible PTEN knockout lines stained for DAPI (blue) and NCAM1 (red). Scale bar indicates 20 μm. (C) Western blot of U2OS cells showing parental line (left) and LMNA knockout lines (right) stained for lamin A protein. (D) Western blot of NGN2-inducible iPS cells showing parental line (left) and PTEN knockout lines (right) stained for PTEN and phospho-AKT protein. Clonal lines C2 and C18 were used for PTEN VIS-seq experiments.

    Article Snippet: pHSG299 (Takara Bio, 3299) pHR-UCOE-SFFV-Zim3-dCas9-P2A-EGFP (Addgene, 188899) piggyFlex (Addgene, 218234) lentiGuide-Puro (Addgene, 52963) pCAG-NLS-HA-Bxb1 (Addgene, 51271) Hyperactive piggyBac transposase expression vector (hyPBase; gift from Jay Shendure lab) pMD2.G (Addgene, 12259) psPAX2 (Addgene, 12260)

    Techniques: In Situ, Sequencing, Cotransfection, Plasmid Preparation, Imaging, Derivative Assay, Knock-Out, Staining, Western Blot