Source 1primary paper2017Neuro-Oncology
Toolkit/LITEs (Light-inducible transcriptional effectors)
LITEs (Light-inducible transcriptional effectors)
Also known as: Light-inducible transcriptional effectors, LITEs
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
LITEs (Light-inducible transcriptional effectors) are a light-controlled, multi-component transcriptional regulation system assembled from two modular components. In the cited study, an optogenetic CRISPR-dCas9 LITE system was used to manipulate endogenous PIM1 transcription in U87 glioblastoma cells in vitro.
Usefulness & Problems
Why this is useful
This system is useful for temporally controlling endogenous gene transcription with light in a targeted CRISPR-dCas9 framework. The cited evidence indicates minute-scale induction or repression, reversibility after light withdrawal, and the potential for graded control with light dose.
Source:
Here we manipulate PIM1 through an optogenetic system using a combination of CRISPR-dCas9 technology and light-inducible heterodimerizing proteins CRY2 and CIB1.
Problem solved
LITEs address the problem of achieving rapid, reversible, and potentially dose-tunable control of endogenous transcription in cultured cells. In the reported application, the system was used to manipulate PIM1 transcription in U87 glioblastoma cells.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
HeterodimerizationTechniques
No technique tags yet.
Target processes
editingtranscriptionInput: Light
Implementation Constraints
The reported implementation used an optogenetic CRISPR-dCas9 LITE system composed of two modular components. Evidence supports use in U87 glioblastoma cells in vitro for endogenous PIM1 transcriptional manipulation, but does not specify guide design, effector domains, or optical hardware parameters.
The supplied evidence is limited to a single in vitro application in U87 glioblastoma cells targeting PIM1. The specific light-responsive protein pair, illumination wavelength, construct architecture, and quantitative performance metrics are not provided in the evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Observations
successMammalian Cell Lineapplication demoU87 cells
Inferred from claim c2 during normalization. In U87 cells, induction or repression of endogenous PIM1 transcription occurred within minutes of light exposure, could theoretically be graded with light dose, and induction was fully reversible after light retraction. Derived from claim c2. Section: abstract. Quoted text: Induction or repression of PIM1 endogenous transcription in U87 cells occurred within minutes of light exposure and the response could be theoretically graded with light dose, with the induction being fully reversible after light retraction.
Source:
light intensity5 mW/cm2light wavelength466 nmmaximum exposure duration24 hresponse time(within minutes)stimulation frequency0.016 Hz
Supporting Sources
Ranked Claims
In U87 cells, induction or repression of endogenous PIM1 transcription occurred within minutes of light exposure, could theoretically be graded with light dose, and induction was fully reversible after light retraction.
Induction or repression of PIM1 endogenous transcription in U87 cells occurred within minutes of light exposure and the response could be theoretically graded with light dose, with the induction being fully reversible after light retraction.
Section: abstract
light intensity 5 mW/cm2light wavelength 466 nmmaximum exposure duration 24 hresponse time within minutesstimulation frequency 0.016 Hz
The study used an optogenetic CRISPR-dCas9 LITE system to manipulate PIM1 transcription in U87 glioblastoma cells in vitro.
Here we manipulate PIM1 through an optogenetic system using a combination of CRISPR-dCas9 technology and light-inducible heterodimerizing proteins CRY2 and CIB1.
Section: abstract
Approval Evidence
1 source2 linked approval claimsfirst-pass slug lites-light-inducible-transcriptional-effectors
LITEs (Light-inducible transcriptional effectors) were used by combining two modular components
Source:
system performancesupports
In U87 cells, induction or repression of endogenous PIM1 transcription occurred within minutes of light exposure, could theoretically be graded with light dose, and induction was fully reversible after light retraction.
Induction or repression of PIM1 endogenous transcription in U87 cells occurred within minutes of light exposure and the response could be theoretically graded with light dose, with the induction being fully reversible after light retraction.
Source:
tool applicationsupports
The study used an optogenetic CRISPR-dCas9 LITE system to manipulate PIM1 transcription in U87 glioblastoma cells in vitro.
Here we manipulate PIM1 through an optogenetic system using a combination of CRISPR-dCas9 technology and light-inducible heterodimerizing proteins CRY2 and CIB1.
Source:
Comparisons
Source-backed strengths
In U87 cells, endogenous PIM1 transcription could be induced or repressed within minutes of light exposure. The response was reported as fully reversible after light retraction, and the study stated that induction could theoretically be graded with light dose.
Ranked Citations
- 1.
Derived from 2 linked claims and 1 validation observations. Example evidence: Induction or repression of PIM1 endogenous transcription in U87 cells occurred within minutes of light exposure and the response could be theoretically graded with light dose, with the induction being fully reversible after light retraction.