A comprehensive library of human transcription factors for cell fate engineering

Abstract

Human pluripotent stem cells (hPSCs) offer an unprecedented opportunity to model diverse cell types and tissues. To enable systematic exploration of the programming landscape mediated by transcription factors (TFs), we present the Human TFome, a comprehensive library containing 1,564 TF genes and 1,732 TF splice-isoforms. By screening the library in three hPSC lines, we discovered 290 TFs, including 241 previously unreported, that induce differentiation in four days without alteration of external soluble or biomechanical cues. We used four of the hits to program hPSCs into neurons, fibroblasts, oligodendrocytes and vascular endothelial–like cells that have molecular and functional similarity to primary cells. Our cell-autonomous approach enabled parallel programming of hPSCs into multiple cell types simultaneously. We also demonstrated orthogonal programming by including oligodendrocyte-inducible hPSCs with unmodified hPSCs to generate cerebral organoids, which expedited in situ myelination. Large-scale combinatorial screening of the Human TFome will complement other strategies for cell engineering based on developmental biology and computational systems biology.

Applications of iPSC-derived beta cells from patients with diabetes

“…These tools could identify iPSC or early-stage markers that define suitability for an efficient SC-islet differentiation 65 or genes to overexpress in iPSCs to initiate or improve differentiation. 91 In addition, reporters to enable robust readouts of function and maturation after cell fate specification would help both cell therapy development and drug screening applications. 92 , 93 , 94 Ideally, SC-islets would be indistinguishable from primary islets for transplantation, specifically in terms of glucose-stimulated insulin secretion, but further protocol improvements are needed to achieve this.…”

Novel Tools and Investigative Approaches for the Study of Oligodendrocyte Precursor Cells (NG2-Glia) in CNS Development and Disease

“…Differentiation of OPCs and subsequently OLs from hiPSCs or hESCs can be achieved by recapitulating the signaling processes associated with differentiation (Goldman and Kuypers, 2015 ), either pharmacologically by timely addition of agonist and antagonists during the differentiation process (Douvaras and Fossati, 2015 ), or by driving expression of key regulatory genes, such as SOX10 and OLIG2 in ESCs (Pawlowski et al, 2017 ), or SOX10, OLIG2 and NKX6.2 (Ehrlich et al, 2017 ) and SOX9 in hiPSCs (Ng et al, 2020 ). A combination of both pharmacological and genetic differentiation methodologies can also be used to great synergistic effect (García-León et al, 2020 ).…”

“…As mentioned, these protocols enable the production of relatively large amounts of OPCs and OLs, which can be used therapeutically (Egawa et al, 2016 ; Morales Pantoja et al, 2020 ) or for experimentation. Xenograft experiments have shown that hiPSC-derived OPCs/OLs survive and can influence disease processes in model animals (Wang et al, 2013 b; Kawabata et al, 2016 ; Feng et al, 2020 ; Ng et al, 2020 ). Finally, and perhaps most illuminating are studies where patient-derived iPSCs are used to make OPCs/OLs, which are then used to study disease processes (for review: Chanoumidou et al, 2020 ).…”

PROGRESS OF ORGANISMS HEALTH & INTELLIGENCE VIA NANITES & REAL-TIME MONITORING OF THEM VIA WIRELESS NETWORKS & MECHATRONIC SYSTEMS

“…II. Implantation for a specific period of time of biodegradable nanobiosensors -nanites -nanomachines -molecular machines for therapeutic issues [3] We can also send [with automation or with technological mandates via ultrasound pulses (ultrasound scan will monitor the nanites -nanomachines -molecular machines for when will arrive to the targets in order to hook up there via activation)] each time when it is a must for health issues, safe biodegradable nanitesnanomachines -molecular machines (from safe biomaterials for organisms without metals in order to there is not a problem with MRI & etc scans) which will release safe therapeutic biochemical -medical substances to the correct -desirable cells [safe biodegradable nanites -nanomachines -molecular machines will invade & bind to the correct -desirable cells because of the analogical specific substances that will have for each occasion to their exterior artificial or/and biochemical membrane (like the membrane of cells) or/and they will invade & bind to the correct -desirable cells through their detectable nanobiosensors] & which will monitor the cause of disease & it's development & later will monitor the actions & interactions of safe therapeutic biochemical -medical substances within the cells before be dissolved (for the understanding of which is the safest way to eliminate each disease through safe therapeutic medical substances) for the confrontation of all type of diseases.…”