Source 1primary paper2023Biology
Toolkit/p21-LINuS
p21-LINuS
Also known as: AsLOV-based light-inducible nuclear localization signal p21 system, LINuS
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
p21-LINuS is a light-controllable p21 construct created by fusing p21 to the AsLOV-based light-inducible nuclear localization signal (LINuS). It was used to optogenetically control p21 subcellular localization and nuclear function, increasing the fraction of cells arrested in G1 phase under blue LED illumination.
Usefulness & Problems
Why this is useful
This system enables optical control of p21 localization and cell-cycle regulation with tunability through blue LED exposure interval and light dose. In producer cell lines, the reported increase in G1-arrested cells correlated with increased cell-specific productivity of secreted alkaline phosphatase, indicating utility for bioprocess optimization.
Source:
Light-controllable p21 implemented in producer cell lines could be applied to steer the uncoupling of cell proliferation and cell cycle arrest at the G1 phase optimizing the production of biotherapeutic proteins.
Source:
Here, we applied an optogenetic approach to control the subcellular localization of p21 and its nuclear functions.
Problem solved
p21-LINuS addresses the problem of dynamically controlling p21 nuclear activity and G1 arrest with light rather than static genetic perturbation. The cited application potential is to uncouple cell proliferation from G1 cell cycle arrest in producer cell lines to optimize biotherapeutic protein production.
Source:
Light-controllable p21 implemented in producer cell lines could be applied to steer the uncoupling of cell proliferation and cell cycle arrest at the G1 phase optimizing the production of biotherapeutic proteins.
Problem links
Need conditional recombination or state switching
Derivedp21-LINuS is a light-controllable p21 construct generated by fusing p21 to the AsLOV-based light-inducible nuclear localization signal (LINuS). It was used to optogenetically control p21 subcellular localization and nuclear function, increasing the fraction of cells arrested in G1 phase under blue LED illumination.
Need inducible protein relocalization or recruitment
Derivedp21-LINuS is a light-controllable p21 construct generated by fusing p21 to the AsLOV-based light-inducible nuclear localization signal (LINuS). It was used to optogenetically control p21 subcellular localization and nuclear function, increasing the fraction of cells arrested in G1 phase under blue LED illumination.
Need precise spatiotemporal control with light input
Derivedp21-LINuS is a light-controllable p21 construct generated by fusing p21 to the AsLOV-based light-inducible nuclear localization signal (LINuS). It was used to optogenetically control p21 subcellular localization and nuclear function, increasing the fraction of cells arrested in G1 phase under blue LED illumination.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
light-inducible nuclear localizationlight-inducible nuclear localizationMembrane Recruitmentoptical control of subcellular localizationoptical control of subcellular localizationTechniques
No technique tags yet.
Target processes
localizationrecombinationInput: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: multi component delivery burdenimplementation constraint: spectral hardware requirementoperating role: reporterswitch architecture: multi componentswitch architecture: recruitment
The tool is implemented as a fusion of p21 with the AsLOV-based light-inducible nuclear localization signal, LINuS. Blue LED illumination was used as the input, and exposure interval and light dose were varied to tune system output; no additional construct architecture, cofactor requirements, or delivery details are provided in the supplied evidence.
The supplied evidence is limited to a single study context and does not provide quantitative performance metrics, kinetics, reversibility, or cell-type breadth for p21-LINuS specifically. Independent replication and validation beyond the reported producer-cell application are not documented in the provided evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Light-controllable p21 in producer cell lines could be applied to uncouple cell proliferation from G1 cell cycle arrest to optimize biotherapeutic protein production.
Light-controllable p21 implemented in producer cell lines could be applied to steer the uncoupling of cell proliferation and cell cycle arrest at the G1 phase optimizing the production of biotherapeutic proteins.
Both p21-CRY2/CIB1 and p21-LINuS increased the amount of cells arrested in the G1 phase.
Both systems, p21-CRY2/CIB1 and p21-LINuS, increased the amounts of cells arrested in the G1 phase
Optogenetically controlled p21 systems were used to control the subcellular localization and nuclear functions of p21.
Here, we applied an optogenetic approach to control the subcellular localization of p21 and its nuclear functions.
The increase in G1-arrested cells correlated with increased cell-specific productivity of secreted alkaline phosphatase.
Both systems, p21-CRY2/CIB1 and p21-LINuS, increased the amounts of cells arrested in the G1 phase correlating with the increased cell-specific productivity of the reporter-protein-secreted alkaline phosphatase.
Blue LED exposure interval and light dose enabled fine-tuning of the optogenetic p21 systems.
Varying the intervals of blue LED light exposure and the light dose enable the fine-tuning of the systems.
Approval Evidence
1 source5 linked approval claimsfirst-pass slug p21-linus
To generate light-controllable p21, appropriate fusions with ... the AsLOV-based light-inducible nuclear localization signal, LINuS, were used. Both systems, p21-CRY2/CIB1 and p21-LINuS...
Source:
application potentialsupports
Light-controllable p21 in producer cell lines could be applied to uncouple cell proliferation from G1 cell cycle arrest to optimize biotherapeutic protein production.
Light-controllable p21 implemented in producer cell lines could be applied to steer the uncoupling of cell proliferation and cell cycle arrest at the G1 phase optimizing the production of biotherapeutic proteins.
Source:
cell cycle arrestsupports
Both p21-CRY2/CIB1 and p21-LINuS increased the amount of cells arrested in the G1 phase.
Both systems, p21-CRY2/CIB1 and p21-LINuS, increased the amounts of cells arrested in the G1 phase
Source:
functional effectsupports
Optogenetically controlled p21 systems were used to control the subcellular localization and nuclear functions of p21.
Here, we applied an optogenetic approach to control the subcellular localization of p21 and its nuclear functions.
Source:
productivity effectsupports
The increase in G1-arrested cells correlated with increased cell-specific productivity of secreted alkaline phosphatase.
Both systems, p21-CRY2/CIB1 and p21-LINuS, increased the amounts of cells arrested in the G1 phase correlating with the increased cell-specific productivity of the reporter-protein-secreted alkaline phosphatase.
Source:
tunabilitysupports
Blue LED exposure interval and light dose enabled fine-tuning of the optogenetic p21 systems.
Varying the intervals of blue LED light exposure and the light dose enable the fine-tuning of the systems.
Source:
Comparisons
Source-backed strengths
The construct was reported to control both subcellular localization and nuclear functions of p21, and it increased the amount of cells arrested in G1 phase. Its response was fine-tunable by adjusting blue LED exposure interval and light dose, and G1 enrichment correlated with higher cell-specific productivity of secreted alkaline phosphatase.
p21-LINuS and CRY2-talin/CIBN-CAAX optogenetic plasma membrane recruitment system address a similar problem space because they share localization, recombination.
Shared frame: same top-level item type; shared target processes: localization, recombination; shared mechanisms: membrane_recruitment; same primary input modality: light
Compared with FUN-LOV
p21-LINuS and FUN-LOV address a similar problem space because they share localization, recombination.
Shared frame: same top-level item type; shared target processes: localization, recombination; shared mechanisms: membrane_recruitment; same primary input modality: light
Relative tradeoffs: appears more independently replicated; looks easier to implement in practice.
Compared with iLID/SspB
p21-LINuS and iLID/SspB address a similar problem space because they share localization, recombination.
Shared frame: same top-level item type; shared target processes: localization, recombination; shared mechanisms: membrane_recruitment; same primary input modality: light
Relative tradeoffs: appears more independently replicated; looks easier to implement in practice.
Ranked Citations
- 1.
Derived from 5 linked claims. Example evidence: Light-controllable p21 implemented in producer cell lines could be applied to steer the uncoupling of cell proliferation and cell cycle arrest at the G1 phase optimizing the production of biotherapeutic proteins.