Events

ISSCR 2018

ISSCR 2018

Meet scientists from the Allen Institute for Cell Science at this year's ISSCR Annual Meeting in Melbourne. Attend our Innovation Showcase, preview our presentations & posters, and keep up with us on social media (#ISSCR2018).

06/20/2018 to 06/23/2018

Innovation Showcase

Allen Institute for Cell Science: Providing Stem Cell & Gene Editing Resources to Empower your Research

Presentation Date & Time: Thursday, June 21, 11:30am-12:30pm
Location: Melbourne Room 1, Level 2

We will share details about our legacy collection of endogenous, fluorescently-tagged hiPS cell lines highlighting key structures within the cell. You will hear how these lines were generated, including gene editing strategies, the plasmids used to generate them and the substantial quality controls performed before making these cells and plasmids publicly available. We will share details about our efforts to differentiate these lines to cardiomyocytes, featuring images from some of our newer ‘cardio-specific’ lines. We will touch briefly on the other resources available on our website (www. allencell.org), including our large, high replicate 3D image data sets showing the subcellular localization of each of our tagged structures, our microscopy pipeline workflow and 3D segmentations. We will also discuss machine learning and label-free approaches to build integrated and predictive models of cell organization, and will wrap up the presentation with a guided tour through our website to familiarize you with where all these resources can be found.

Presentation

Concurrent IIA: Gene Editing
Session Date & Time: Thursday, June 21, 4:00-6:00pm
Location: Melbourne Room 1, Level 2
Speaker: Ruwanthi Gunawardane, 4:30-4:45pm, "Endogenous Gene Tagging with CRISPR/Cas9 to Illuminate Cell Organization and Dynamics"

The Allen Institute for Cell Science is creating a dynamic visual model of hiPSC organization to understand and predict normal and pathological cell states. Towards this goal, we have created the “Allen Cell Collection”; an open source collection of fluorescently tagged human induced pluripotent stem cell (hiPSC) lines representing the major organelles of the cell. Our approach utilizes CRISPR/Cas9 to introduce fluorescent tags via homology driven repair (HDR) into the genomic locus of interest. Editing yields isogenic hiPSC lines expressing fusion proteins unique to each cell line under endogenous regulation. Live cell imaging, image analysis, computational modeling, and open distribution of tools and data to the scientific community define our endeavor. Here we present the CRISPCR/Cas9- based gene editing strategy and workflow used to generate ~25 GFP-tagged clonal hiPSC lines representing the major organelles of the cell including the nucleus, mitochondria, adhesions, cytoskeleton, golgi and ER. We have also developed related editing methods for introducing red fluorescent proteins, tagging dual structures, safe harbor edits, and differentiation specific gene tagging. We will describe our genotyping strategy to identify clones harboring precisely incorporated FP tags including off-target and NGS analysis and present trends observed during precise editing. Clonal lines also undergo various quality control assays for ensuring genomic stability, pluripotency, and confirmation of subcellular localization prior to use in live cell imaging and distribution. We will highlight the utility of these cell lines for generating image-based integrated models of cell organization and dynamics and discuss the potential applications of these endogenously tagged lines for basic science and disease modeling.

Posters

Wednesday, June 20

W-3031 Developing a Visual Model and Genomic Map for Cardiomyocyte Differentiation
Poster Session 1-ODD, 6:30-7:30pm

Thursday, June 21

T-1085 Integrated Cytoplasmic Reorganization During Human Inducible Pluripotent Stem Cell Mitosis
Poster Session 2-EVEN, 7:00-8:00pm

ISSCR Annual Meeting

Melbourne Convention & Exhibition Centre
1 Convention Centre Place
South Wharf VIC 3006
Melbourne, Australia

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