From 51c24316f6ddda4a5999d16beca5dc48626de90d Mon Sep 17 00:00:00 2001 From: "sara.carlson" Date: Fri, 31 Jul 2026 05:24:03 +0000 Subject: [PATCH] =?UTF-8?q?Update=20Cell=20Line=20=E2=80=9CAICS-80-69/inde?= =?UTF-8?q?x=E2=80=9D?= MIME-Version: 1.0 Content-Type: text/plain; charset=UTF-8 Content-Transfer-Encoding: 8bit --- src/pages/cell-line/AICS-80-69/index.md | 134 +++++++++++++++--------- 1 file changed, 87 insertions(+), 47 deletions(-) diff --git a/src/pages/cell-line/AICS-80-69/index.md b/src/pages/cell-line/AICS-80-69/index.md index 49d1778c..954eed64 100644 --- a/src/pages/cell-line/AICS-80-69/index.md +++ b/src/pages/cell-line/AICS-80-69/index.md @@ -2,6 +2,7 @@ templateKey: cell-line cell_line_id: 80 status: data complete +date: 2026-07-30T22:24:00.000Z clone_number: 69 parental_line: 0 genetic_modifications: @@ -12,18 +13,35 @@ genetic_modifications: order_link: https://www.coriell.org/0/Sections/Search/Sample_Detail.aspx?Ref=AICS-0080-069&PgId=166 certificate_of_analysis: https://www.coriell.org/0/PDF/Allen/ipsc/AICS-0080-069_CofA.pdf donor_plasmid: https://www.addgene.org/114410/ -hpscreg_certificate_link: https://hpscreg.eu/cell-line/UCSFi001-A-35 images_and_videos: images: - image: single_plane_image_cl69.jpg - caption: "Single, mid-level plane of cells in a live hiPS cell colony expressing mEGFP-tagged RNA-binding protein FUS in control cells (left panel) and in the presence of 500 µM sodium arsenite for 15 min (right panel). Cells were imaged in 3D on a spinning-disk confocal microscope. Scale bar, 5 µm. " + caption: "Single, mid-level plane of cells in a live hiPS cell colony expressing + mEGFP-tagged RNA-binding protein FUS in control cells (left panel) and + in the presence of 500 µM sodium arsenite for 15 min (right panel). + Cells were imaged in 3D on a spinning-disk confocal microscope. Scale + bar, 5 µm. " - image: Main_cell_line_morphology.jpg - caption: "Viability and colony formation photographed 3 days post-thaw at 4X magnification. Cells were treated with ROCK inhibitor for 24 hrs post-thaw." + caption: Viability and colony formation photographed 3 days post-thaw at 4X + magnification. Cells were treated with ROCK inhibitor for 24 hrs + post-thaw. + - image: Western blot documentation FUS final clone only_final.jpg + - image: final FUS IF 110918.jpg videos: - video: https://player.vimeo.com/video/307598559 - caption: "Z-stack of a live hiPS cell colony expressing mEGFP-tagged RNA-binding protein FUS in control cells (left panel) and in the presence of 500 µM sodium arsenite for 15 min (right panel). Cells were imaged in 3D on a spinning-disk confocal microscope. Movie starts at the bottom of the cells and ends at the top. Scale bar, 5µm." + caption: Z-stack of a live hiPS cell colony expressing mEGFP-tagged RNA-binding + protein FUS in control cells (left panel) and in the presence of 500 µM + sodium arsenite for 15 min (right panel). Cells were imaged in 3D on a + spinning-disk confocal microscope. Movie starts at the bottom of the + cells and ends at the top. Scale bar, 5µm. - video: https://player.vimeo.com/video/307598542 - caption: "Time-lapse movie of a live hiPS cell colony expressing mEGFP-tagged RNA-binding protein FUS. Six minutes after the introduction of 500 µM sodium arsenite, cells were imaged every 5 sec in 3D on a spinning-disk confocal microscope. A single mid-level plane is shown. The inset is a 2.5x enlargement of the boxed region to show detail of aggregate formation. Frames were histogram matched to adjust for photobleaching. Movie plays at 25x real time. Scale bar, 5 µm." + caption: Time-lapse movie of a live hiPS cell colony expressing mEGFP-tagged + RNA-binding protein FUS. Six minutes after the introduction of 500 µM + sodium arsenite, cells were imaged every 5 sec in 3D on a spinning-disk + confocal microscope. A single mid-level plane is shown. The inset is a + 2.5x enlargement of the boxed region to show detail of aggregate + formation. Frames were histogram matched to adjust for photobleaching. + Movie plays at 25x real time. Scale bar, 5 µm. editing_design: ncbi_isoforms: - n @@ -31,75 +49,97 @@ editing_design: linker: ENLYFQGAAKFKETAAAKFERQHMDSGGGGSSGPSGSSSLEVLFQGPLSSSGPSGS cas9: Wildtype spCas9 diagrams: - - title: "mEGFP Insert" + - title: mEGFP Insert images: - image: EditingDesign_gene_figure.png - caption: "Top: FUS locus showing 3 FUS isoforms; Bottom: Zoom in on mEGFP insertion site at FUS C-terminal exon" -category_labels: - - Stress - - Nuclear Structure + caption: "Top: FUS locus showing 3 FUS isoforms; Bottom: Zoom in on mEGFP + insertion site at FUS C-terminal exon" genomic_characterization: diagrams: - - title: "Schematic of Junctions" + - title: Schematic of Junctions images: - image: /img/shared/GenomicCharacterization_junction_schematic_mEGFP.png - - title: "Karyotype Analysis" + - title: Karyotype Analysis images: - image: KARYO_cl69_AICS_80_FUS_mEGFP_cl.jpg - caption: "After cells banks were created, one vial was thawed and 30 G-banded metaphase cells were karyotyped." + caption: After cells banks were created, one vial was thawed and 30 G-banded + metaphase cells were karyotyped. amplified_junctions: - - edited_gene: "FUS-mEGFP" - junction: "5'" + - edited_gene: FUS-mEGFP + junction: 5' expected_size: "2178" - confirmed_sequence: "Pass" - - edited_gene: "FUS-mEGFP" - junction: "3'" + confirmed_sequence: Pass + - edited_gene: FUS-mEGFP + junction: 3' expected_size: "1485" - confirmed_sequence: "Pass" - - edited_gene: "FUS-mEGFP" - junction: "WT internal" + confirmed_sequence: Pass + - edited_gene: FUS-mEGFP + junction: WT internal expected_size: "694" - confirmed_sequence: "Pass" - - edited_gene: "FUS-mEGFP" - junction: "Full junctional allele" + confirmed_sequence: Pass + - edited_gene: FUS-mEGFP + junction: Full junctional allele expected_size: "Wild Type: 2722 bp; Insert: 3715 bp" - confirmed_sequence: "Pass" - junction_table_caption: "PCR amplified 5', 3', WT, and full allele junctions. 5', 3', and WT junctions were Sanger sequenced to check for precise mEGFP insertion. Primers were designed to exclude amplification from the donor plasmid." + confirmed_sequence: Pass + junction_table_caption: PCR amplified 5', 3', WT, and full allele junctions. 5', + 3', and WT junctions were Sanger sequenced to check for precise mEGFP + insertion. Primers were designed to exclude amplification from the donor + plasmid. ddpcr: - tag: FUS-mEGFP clone: 69 fp_ratio: 0.52 - plasmid: 0.0 - ddpcr_caption: "FP:RPP30 ratio from ddPCR assay; values = 0.5 +/- 0.1 indicate heterozygous clone, values = 1 +/- 0.1 indicate homozygous clone. Plasmid: AmpR/RPP30 ratio from ddPCR assay; values <0.1 indicate clone with no detectable plasmid integration. RPP30 is known 2n reference gene." + plasmid: 0 + ddpcr_caption: "FP:RPP30 ratio from ddPCR assay; values = 0.5 +/- 0.1 indicate + heterozygous clone, values = 1 +/- 0.1 indicate homozygous clone. Plasmid: + AmpR/RPP30 ratio from ddPCR assay; values <0.1 indicate clone with no + detectable plasmid integration. RPP30 is known 2n reference gene." +category_labels: + - Stress + - Nuclear Structure +hpscreg_certificate_link: https://hpscreg.eu/cell-line/UCSFi001-A-35 stem_cell_characteristics: pluripotency_analysis: - - marker: "NANOG" + - marker: NANOG positive_cells: 9.91 - - marker: "SOX2" + - marker: SOX2 positive_cells: 99.94 - - marker: "OCT4" + - marker: OCT4 positive_cells: 99.54 - - marker: "SSEA-1" + - marker: SSEA-1 positive_cells: 0.45 - - marker: "SSEA-3" + - marker: SSEA-3 positive_cells: null - - marker: "TRA-160" + - marker: TRA-160 positive_cells: 99.92 - pluripotency_caption: "iPSCs were stained with directly conjugated antibodies from BD Biosciences, acquired using a FACSAria III Fusion (BD Biosciences), and analyzed using FlowJo software (Treestar, Inc.). Doublets were excluded, then marker-specific gates were set according to corresponding fluorescence-minus-one (FMO) controls." + pluripotency_caption: iPSCs were stained with directly conjugated antibodies + from BD Biosciences, acquired using a FACSAria III Fusion (BD Biosciences), + and analyzed using FlowJo software (Treestar, Inc.). Doublets were excluded, + then marker-specific gates were set according to corresponding + fluorescence-minus-one (FMO) controls. trilineage_differentiation: - - germ_layer: "Ectoderm" - marker: "PAX6" + - germ_layer: Ectoderm + marker: PAX6 percent_positive_cells: Pass - - germ_layer: "Endoderm" - marker: "SOX17" + - germ_layer: Endoderm + marker: SOX17 percent_positive_cells: Pass - - germ_layer: "Mesoderm" - marker: "Brachyury" + - germ_layer: Mesoderm + marker: Brachyury percent_positive_cells: Pass - trilineage_caption: "iPSCs were subjected to a 5-7 day, non-terminal, directed differentiation using the STEMdiff™ Trilineage Differentiation Kit (STEMCELL Technologies, Inc.). Total RNA was isolated from each lineage specific differentiation and assayed via ddPCR for the expression of lineage specific transcripts; Pax6(Ectoderm), Sox17(Endoderm) and Brachyury(Mesoderm)." + trilineage_caption: iPSCs were subjected to a 5-7 day, non-terminal, directed + differentiation using the STEMdiff™ Trilineage Differentiation Kit (STEMCELL + Technologies, Inc.). Total RNA was isolated from each lineage specific + differentiation and assayed via ddPCR for the expression of lineage specific + transcripts; Pax6(Ectoderm), Sox17(Endoderm) and Brachyury(Mesoderm). cardiomyocyte_differentiation: - troponin_percent_positive: "79.5-82.5 (2)" - day_of_beating_percent: "100 (3)" - day_of_beating_range: "d8-d9" - cardiomyocyte_differentiation_caption: "iPSCs were differentiated to cardiomyocytes and observed for initiation of beating starting at day 6. At ~day 12, cells were fixed and stained with anti-cardiac Troponin T (BD Biosciences) and gating was based on an isotype control. Ranges observed across multiple experiments are shown for Troponin T and Day of beating initiation; number of experiments is shown in (). " ---- \ No newline at end of file + troponin_percent_positive: 79.5-82.5 (2) + day_of_beating_percent: 100 (3) + day_of_beating_range: d8-d9 + cardiomyocyte_differentiation_caption: "iPSCs were differentiated to + cardiomyocytes and observed for initiation of beating starting at day 6. At + ~day 12, cells were fixed and stained with anti-cardiac Troponin T (BD + Biosciences) and gating was based on an isotype control. Ranges observed + across multiple experiments are shown for Troponin T and Day of beating + initiation; number of experiments is shown in (). " +---