TY - JOUR TI - Spatiotemporal mosaic self-patterning of pluripotent stem cells using CRISPR interference AU - Libby, Ashley RG AU - Joy, David A AU - So, Po-Lin AU - Mandegar, Mohammad A AU - Muncie, Jonathon M AU - Mendoza-Camacho, Federico N AU - Weaver, Valerie M AU - Conklin, Bruce R AU - McDevitt, Todd C VL - 7 PY - 2018 DA - 2018/10/09 SP - e36045 C1 - eLife 2018;7:e36045 DO - 10.7554/eLife.36045 UR - https://doi.org/10.7554/eLife.36045 AB - Morphogenesis involves interactions of asymmetric cell populations to form complex multicellular patterns and structures comprised of distinct cell types. However, current methods to model morphogenic events lack control over cell-type co-emergence and offer little capability to selectively perturb specific cell subpopulations. Our in vitro system interrogates cell-cell interactions and multicellular organization within human induced pluripotent stem cell (hiPSC) colonies. We examined effects of induced mosaic knockdown of molecular regulators of cortical tension (ROCK1) and cell-cell adhesion (CDH1) with CRISPR interference. Mosaic knockdown of ROCK1 or CDH1 resulted in differential patterning within hiPSC colonies due to cellular self-organization, while retaining an epithelial pluripotent phenotype. Knockdown induction stimulates a transient wave of differential gene expression within the mixed populations that stabilized in coordination with observed self-organization. Mosaic patterning enables genetic interrogation of emergent multicellular properties, which can facilitate better understanding of the molecular pathways that regulate symmetry-breaking during morphogenesis. KW - pluripotent stem cells KW - morphogenesis KW - bio-engineering JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -