Toolkit/NanoLuc luciferase reporter assay

NanoLuc luciferase reporter assay

Assay Method·Research·Since 2017

Also known as: NanoLuc® luciferase reporter assay

Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.

Summary

The NanoLuc luciferase reporter assay is a bioluminescent functional reporter assay used to test guide RNA efficiency. In the cited study, it served as a pre-screening step to identify optimal guide RNAs before downstream cell-based experiments with an optogenetic CRISPR-dCas9 LITE system.

Usefulness & Problems

Why this is useful

This assay is useful for functionally ranking guide RNAs before more complex cellular experiments. The cited evidence supports its use as an upstream screening method to select guides with optimal efficiency for subsequent genome-engineering workflows.

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

It addresses the need to identify efficient guide RNAs prior to downstream cell-based experiments. In the cited application, it reduced uncertainty in guide selection before using optogenetic CRISPR-dCas9 LITE to manipulate transcription in U87 glioblastoma cells.

Taxonomy & Function

Primary hierarchy

Technique Branch

Method: A concrete measurement method used to characterize an engineered system.

Target processes

No target processes tagged yet.

Implementation Constraints

cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: multi component delivery burdenoperating role: sensorswitch architecture: multi componentswitch architecture: recruitment

The evidence indicates that guide RNAs were tested for optimal efficiencies using the NanoLuc luciferase reporter assay before downstream cell-based experiments. No additional implementation details are provided here regarding construct architecture, substrate chemistry, instrumentation, or expression system requirements.

The supplied evidence does not report assay sensitivity, specificity, signal-to-background, throughput, or compatibility across multiple cell types. Validation is limited to a single cited use case as a guide-screening step, with no independent replication provided here.

Validation

Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication

Supporting Sources

Ranked Claims

Claim 1assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 2assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 3assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 4assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 5assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 6assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 7assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 8assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 9assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 10assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 11assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 12assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 13assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 14assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 15assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 16assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 17assay usesupports2017Source 1needs review

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.
Section: abstract
Claim 18system performancesupports2017Source 1needs review

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
Claim 19system performancesupports2017Source 1needs review

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
Claim 20system performancesupports2017Source 1needs review

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
Claim 21system performancesupports2017Source 1needs review

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
Claim 22system performancesupports2017Source 1needs review

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
Claim 23system performancesupports2017Source 1needs review

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
Claim 24system performancesupports2017Source 1needs review

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
Claim 25system performancesupports2017Source 1needs review

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
Claim 26system performancesupports2017Source 1needs review

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
Claim 27system performancesupports2017Source 1needs review

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
Claim 28tool applicationsupports2017Source 1needs review

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
Claim 29tool applicationsupports2017Source 1needs review

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
Claim 30tool applicationsupports2017Source 1needs review

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
Claim 31tool applicationsupports2017Source 1needs review

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
Claim 32tool applicationsupports2017Source 1needs review

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
Claim 33tool applicationsupports2017Source 1needs review

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
Claim 34tool applicationsupports2017Source 1needs review

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
Claim 35tool applicationsupports2017Source 1needs review

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
Claim 36tool applicationsupports2017Source 1needs review

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
Claim 37tool applicationsupports2017Source 1needs review

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 source1 linked approval claimfirst-pass slug nanoluc-luciferase-reporter-assay
Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay

Source:

assay usesupports

Guide RNAs were tested for efficiency using a NanoLuc luciferase reporter assay before downstream cell-based experiments.

Guide RNAs were tested for optimal efficiencies using the NanoLuc® luciferase reporter assay.

Source:

Comparisons

Source-backed strengths

The main demonstrated strength is its use as a functional pre-screen for guide RNA efficiency. The available evidence directly supports this screening role, but does not provide quantitative performance metrics, dynamic range, or comparative benchmarking.

Compared with Field-domain rapid-scan EPR at 240 GHz

NanoLuc luciferase reporter assay and Field-domain rapid-scan EPR at 240 GHz address a similar problem space.

Shared frame: same top-level item type

Compared with fluorescence line narrowing

NanoLuc luciferase reporter assay and fluorescence line narrowing address a similar problem space.

Shared frame: same top-level item type

Compared with native green gel system

NanoLuc luciferase reporter assay and native green gel system address a similar problem space.

Shared frame: same top-level item type

Strengths here: looks easier to implement in practice.

Ranked Citations

  1. 1.
    StructuralSource 1Neuro-Oncology2017Claim 12Claim 11Claim 11

    Extracted from this source document.