Cell Discovery, Год журнала: 2024, Номер 10(1)
Опубликована: Окт. 15, 2024
Язык: Английский
Cell Systems, Год журнала: 2022, Номер 13(12), С. 950 - 973
Опубликована: Дек. 1, 2022
To elucidate principles operating in native biological systems and to develop novel biotechnologies, synthetic biology aims build integrate gene circuits within transcriptional networks. The utility of for cell engineering relies on the ability control expression all constituent transgene components. Transgene silencing, defined as loss over time, persists an obstacle primary cells stem with transgenic cargos. In this review, we highlight challenge that silencing poses robust mammalian cells, outline potential molecular mechanisms present approaches preventing silencing. We conclude a perspective identifying future research directions improving performance circuits.
Язык: Английский
Процитировано
97
Nature Reviews Genetics, Год журнала: 2024, Номер 25(6), С. 381 - 400
Опубликована: Янв. 18, 2024
Язык: Английский
Процитировано
63
Nature Reviews Rheumatology, Год журнала: 2024, Номер 20(2), С. 81 - 100
Опубликована: Янв. 22, 2024
Язык: Английский
Процитировано
30
Cell stem cell, Год журнала: 2023, Номер 30(1), С. 10 - 19
Опубликована: Янв. 1, 2023
Язык: Английский
Процитировано
33
Development, Год журнала: 2024, Номер 151(1)
Опубликована: Янв. 1, 2024
ABSTRACT Cell-cell interactions are central to development, but exploring how a change in any given cell relates changes the neighbour of that can be technically challenging. Here, we review recent developments synthetic biology and image analysis helping overcome this problem. We highlight opportunities presented by these advances discuss limitations applying them developmental model systems.
Язык: Английский
Процитировано
10
The EMBO Journal, Год журнала: 2024, Номер 43(18), С. 4110 - 4135
Опубликована: Июль 12, 2024
Cell communication coordinates developmental processes, maintains homeostasis, and contributes to disease. Therefore, understanding the relationship between cells in a shared environment is crucial. Here we introduce Positive Ultra-bright Fluorescent Fusion For Identifying Neighbours (PUFFFIN), cell neighbour-labelling system based upon secretion uptake of positively supercharged fluorescent protein s36GFP. We fused s36GFP mNeonGreen or HaloTag, facilitating ultra-bright, sensitive, colour-of-choice labelling. Secretor transfer PUFFFIN neighbours while retaining nuclear mCherry, making identification, isolation, investigation live straightforward. can be delivered cells, tissues, embryos on customisable single-plasmid construct composed interchangeable components with option incorporate any transgene. This versatility enables manipulation properties, simultaneously labelling surrounding culture vivo. use ask whether pluripotent adjust pace differentiation synchronise their during exit from naïve pluripotency. offers simple, approach profile non-cell-autonomous responses natural induced changes identity behaviour.
Язык: Английский
Процитировано
5
EMBO Reports, Год журнала: 2024, Номер unknown
Опубликована: Сен. 27, 2024
Abstract Morphogens, locally produced signaling molecules, form a concentration gradient to guide tissue patterning. Tissue patterns emerge as collaboration between morphogen diffusion and responsive cell behaviors, but the mechanisms through which diffusing morphogens define precise spatial amidst biological fluctuations remain unclear. To investigate how cells respond proteins generate patterns, we develop SYMPLE3D, 3D culture platform. By engineering gene expression artificial morphogens, observe that coupling signals with cadherin-based adhesion is sufficient convert into distinct domains. Morphogen-induced cadherins gather activated single domain, removing ectopically cells. In addition, reveal switch-like induction of cadherin-mediated compaction mixing, homogenizing within uniformly domain sharp boundary. These findings highlight cooperation gradients in robust patterning introduce novel method for new domains organoids.
Язык: Английский
Процитировано
3
Biology Open, Год журнала: 2025, Номер 14(1)
Опубликована: Янв. 15, 2025
ABSTRACT In the developing mouse ventral spinal cord, HES5, a transcription factor downstream of Notch signalling, is expressed as evenly spaced clusters high HES5-expressing neural progenitor cells along dorsoventral axis. While signalling requires direct membrane contact for its activation, we have previously shown mathematically that needs to extend beyond neighbouring HES5 pattern emerge. However, presence cellular structures could enable such long-distance was unclear. Here, report protrusions are present all apicobasal axis individual cells. Through live imaging, show these dynamically and retract reaching lengths up ∼20 µm, enough adjacent The ligand DLL1 found colocalise with protrusions, further supporting idea can be transduced at distance. effect on tested by reducing density using CDC42 inhibitor ML141, leading tendency decrease distance between cell clusters. this not significant leaves an open question about their role in fine-grained organisation neurogenesis.
Язык: Английский
Процитировано
0
ACS Synthetic Biology, Год журнала: 2025, Номер unknown
Опубликована: Май 6, 2025
Historically, studying the development of brain and central nervous system (CNS) tissues has been challenging. Human pluripotent stem cell (hPSC) technology allowed for in vitro reconstitution relevant, early trajectories by using small molecules recombinant proteins to guide differentiation cells toward relevant CNS phenotypes. However, many these protocols fail recapitulate cell-guided programs intrinsic embryonic development, particularly signaling centers that emerge within neural tube during formation. Located on ventral end tube, floor plate acts as one such center pattern dorsal/ventral axis secreting morphogen Sonic Hedgehog (SHH). Here, we present a method synthetic Notch (synNotch) receptor platform regulate SHH production subsequent fate specification. We show widely used configuration orthogonal synNotch ligand green fluorescent protein (GFP) mounted platelet-derived growth factor receptor-β transmembrane chassis does not allow robust artificial synNotch-hPSCs ("receivers") cocultured with ligand-presenting hPSCs ("senders"). discovered refined designs membrane-bound GFP-ligand efficient activation hPSC receivers. A variant this enhanced drives sender:hPSC receiver cocultures gives rise plate-like types seen development. This revised potential morphogenesis studies designed uncover key paradigms human
Язык: Английский
Процитировано
0
bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown
Опубликована: Март 12, 2024
Abstract Synthetic developmental biology uses engineering approaches to understand multicellularity with goals ranging from recapitulating development building synthetic organisms. Current include multicellular patterning, controlling differentiation, and implementing cooperative cellular behaviors in model systems. tools enable these pursuits genetic circuits that drive customized responses arbitrary stimuli, receptors orthogonal signaling channels, light- or drug-inducible systems precise spatial temporal control of cell function. Mouse embryonic stem cells (mESCs) offer a well-studied genetically tractable pluripotent chassis for pursuing questions however, there is minimal characterization existing mESCs we lack toolkits rapid iterative workflows. Here, began address this challenge by characterizing small molecule contact-inducible gene expression differentiation mESCs. We show cell-contact inducible work reliably efficiently payloads. Furthermore, can direct into neurons. Each readily be used on their own combination, opening many possibilities studying principles high precision.
Язык: Английский
Процитировано
2