Toolkit/LUNAS

LUNAS

Multi-Component Switch·Research·Since 2023

Also known as: bioluminescent nucleic acid sensor, RPA-LUNAS, RT-RPA-LUNAS assay

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

Summary

LUNAS is a bioluminescent nucleic acid sensor in which a target double-stranded DNA sequence is recognized by two dCas9-based probes that mediate split NanoLuc luciferase complementation. Reported implementations couple this sensor to recombinase polymerase amplification, including RT-RPA-LUNAS for SARS-CoV-2 RNA detection.

Usefulness & Problems

Why this is useful

LUNAS provides sequence-specific nucleic acid detection with a bioluminescent output that can be monitored in real time. In reported one-pot RPA-coupled formats, it enables rapid, sensitive detection and a ratiometric readout that can be captured with a simple digital camera.

Source:

Here, we developed a bioluminescent nucleic acid sensor (LUNAS) platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.

Source:

showing that RPA-LUNAS is attractive for point-of-care infectious disease testing.

Problem solved

This tool addresses the need for rapid nucleic acid diagnostics that combine sequence-specific recognition with simple optical readout under isothermal conditions. The reported system specifically supports detection of amplified target nucleic acids, including SARS-CoV-2 RNA after RT-RPA.

Source:

showing that RPA-LUNAS is attractive for point-of-care infectious disease testing.

Taxonomy & Function

Primary hierarchy

Mechanism Branch

Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.

Target processes

diagnosticrecombination

Implementation Constraints

The core construct uses a pair of dCas9-based probes designed to bind a target dsDNA sequence and drive split NanoLuc complementation. Reported implementations integrate the sensor with recombinase polymerase amplification in a one-pot assay, and a calibrator luciferase is included for ratiometric luminescence readout and real-time monitoring.

The supplied evidence is limited to a single 2023 study and focuses on RPA-coupled diagnostic use cases, especially SARS-CoV-2. The provided evidence does not describe broader organismal validation, comparative benchmarking against other diagnostic platforms, or independent replication.

Validation

Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication

Supporting Sources

Ranked Claims

Claim 1diagnostic performancesupports2023Source 1needs review

RPA-LUNAS detected SARS-CoV-2 from patient nasopharyngeal swab samples with viral RNA loads of approximately 200 cp/μL within approximately 20 minutes.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation and demonstrated its diagnostic performance for COVID-19 patient nasopharyngeal swab samples. Detection of SARS-CoV-2 from samples with viral RNA loads of ∼200 cp/μL was achieved within ∼20 min
time to detection ∼20 minviral RNA load ∼200 cp/μL
Claim 2diagnostic performancesupports2023Source 1needs review

RPA-LUNAS detected SARS-CoV-2 from patient nasopharyngeal swab samples with viral RNA loads of approximately 200 cp/μL within approximately 20 minutes.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation and demonstrated its diagnostic performance for COVID-19 patient nasopharyngeal swab samples. Detection of SARS-CoV-2 from samples with viral RNA loads of ∼200 cp/μL was achieved within ∼20 min
time to detection ∼20 minviral RNA load ∼200 cp/μL
Claim 3diagnostic performancesupports2023Source 1needs review

RPA-LUNAS detected SARS-CoV-2 from patient nasopharyngeal swab samples with viral RNA loads of approximately 200 cp/μL within approximately 20 minutes.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation and demonstrated its diagnostic performance for COVID-19 patient nasopharyngeal swab samples. Detection of SARS-CoV-2 from samples with viral RNA loads of ∼200 cp/μL was achieved within ∼20 min
time to detection ∼20 minviral RNA load ∼200 cp/μL
Claim 4diagnostic performancesupports2023Source 1needs review

RPA-LUNAS detected SARS-CoV-2 from patient nasopharyngeal swab samples with viral RNA loads of approximately 200 cp/μL within approximately 20 minutes.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation and demonstrated its diagnostic performance for COVID-19 patient nasopharyngeal swab samples. Detection of SARS-CoV-2 from samples with viral RNA loads of ∼200 cp/μL was achieved within ∼20 min
time to detection ∼20 minviral RNA load ∼200 cp/μL
Claim 5diagnostic performancesupports2023Source 1needs review

RPA-LUNAS detected SARS-CoV-2 from patient nasopharyngeal swab samples with viral RNA loads of approximately 200 cp/μL within approximately 20 minutes.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation and demonstrated its diagnostic performance for COVID-19 patient nasopharyngeal swab samples. Detection of SARS-CoV-2 from samples with viral RNA loads of ∼200 cp/μL was achieved within ∼20 min
time to detection ∼20 minviral RNA load ∼200 cp/μL
Claim 6diagnostic performancesupports2023Source 1needs review

RPA-LUNAS detected SARS-CoV-2 from patient nasopharyngeal swab samples with viral RNA loads of approximately 200 cp/μL within approximately 20 minutes.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation and demonstrated its diagnostic performance for COVID-19 patient nasopharyngeal swab samples. Detection of SARS-CoV-2 from samples with viral RNA loads of ∼200 cp/μL was achieved within ∼20 min
time to detection ∼20 minviral RNA load ∼200 cp/μL
Claim 7diagnostic performancesupports2023Source 1needs review

RPA-LUNAS detected SARS-CoV-2 from patient nasopharyngeal swab samples with viral RNA loads of approximately 200 cp/μL within approximately 20 minutes.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation and demonstrated its diagnostic performance for COVID-19 patient nasopharyngeal swab samples. Detection of SARS-CoV-2 from samples with viral RNA loads of ∼200 cp/μL was achieved within ∼20 min
time to detection ∼20 minviral RNA load ∼200 cp/μL
Claim 8integration capabilitysupports2023Source 1needs review

LUNAS can be integrated with recombinase polymerase amplification in a rapid one-pot assay with attomolar sensitivity.

LUNAS is easily integrated with recombinase polymerase amplification (RPA), providing attomolar sensitivity in a rapid one-pot assay.
sensitivity attomolar
Claim 9integration capabilitysupports2023Source 1needs review

LUNAS can be integrated with recombinase polymerase amplification in a rapid one-pot assay with attomolar sensitivity.

LUNAS is easily integrated with recombinase polymerase amplification (RPA), providing attomolar sensitivity in a rapid one-pot assay.
sensitivity attomolar
Claim 10integration capabilitysupports2023Source 1needs review

LUNAS can be integrated with recombinase polymerase amplification in a rapid one-pot assay with attomolar sensitivity.

LUNAS is easily integrated with recombinase polymerase amplification (RPA), providing attomolar sensitivity in a rapid one-pot assay.
sensitivity attomolar
Claim 11integration capabilitysupports2023Source 1needs review

LUNAS can be integrated with recombinase polymerase amplification in a rapid one-pot assay with attomolar sensitivity.

LUNAS is easily integrated with recombinase polymerase amplification (RPA), providing attomolar sensitivity in a rapid one-pot assay.
sensitivity attomolar
Claim 12integration capabilitysupports2023Source 1needs review

LUNAS can be integrated with recombinase polymerase amplification in a rapid one-pot assay with attomolar sensitivity.

LUNAS is easily integrated with recombinase polymerase amplification (RPA), providing attomolar sensitivity in a rapid one-pot assay.
sensitivity attomolar
Claim 13integration capabilitysupports2023Source 1needs review

LUNAS can be integrated with recombinase polymerase amplification in a rapid one-pot assay with attomolar sensitivity.

LUNAS is easily integrated with recombinase polymerase amplification (RPA), providing attomolar sensitivity in a rapid one-pot assay.
sensitivity attomolar
Claim 14integration capabilitysupports2023Source 1needs review

LUNAS can be integrated with recombinase polymerase amplification in a rapid one-pot assay with attomolar sensitivity.

LUNAS is easily integrated with recombinase polymerase amplification (RPA), providing attomolar sensitivity in a rapid one-pot assay.
sensitivity attomolar
Claim 15readout capabilitysupports2023Source 1needs review

A calibrator luciferase enables robust ratiometric readout and real-time monitoring of the RPA reaction using a simple digital camera.

A calibrator luciferase is included for a robust ratiometric readout, enabling real-time monitoring of the RPA reaction using a simple digital camera.
Claim 16readout capabilitysupports2023Source 1needs review

A calibrator luciferase enables robust ratiometric readout and real-time monitoring of the RPA reaction using a simple digital camera.

A calibrator luciferase is included for a robust ratiometric readout, enabling real-time monitoring of the RPA reaction using a simple digital camera.
Claim 17readout capabilitysupports2023Source 1needs review

A calibrator luciferase enables robust ratiometric readout and real-time monitoring of the RPA reaction using a simple digital camera.

A calibrator luciferase is included for a robust ratiometric readout, enabling real-time monitoring of the RPA reaction using a simple digital camera.
Claim 18readout capabilitysupports2023Source 1needs review

A calibrator luciferase enables robust ratiometric readout and real-time monitoring of the RPA reaction using a simple digital camera.

A calibrator luciferase is included for a robust ratiometric readout, enabling real-time monitoring of the RPA reaction using a simple digital camera.
Claim 19readout capabilitysupports2023Source 1needs review

A calibrator luciferase enables robust ratiometric readout and real-time monitoring of the RPA reaction using a simple digital camera.

A calibrator luciferase is included for a robust ratiometric readout, enabling real-time monitoring of the RPA reaction using a simple digital camera.
Claim 20readout capabilitysupports2023Source 1needs review

A calibrator luciferase enables robust ratiometric readout and real-time monitoring of the RPA reaction using a simple digital camera.

A calibrator luciferase is included for a robust ratiometric readout, enabling real-time monitoring of the RPA reaction using a simple digital camera.
Claim 21readout capabilitysupports2023Source 1needs review

A calibrator luciferase enables robust ratiometric readout and real-time monitoring of the RPA reaction using a simple digital camera.

A calibrator luciferase is included for a robust ratiometric readout, enabling real-time monitoring of the RPA reaction using a simple digital camera.
Claim 22sample processing capabilitysupports2023Source 1needs review

The RT-RPA-LUNAS assay allows SARS-CoV-2 RNA detection without the need for RNA isolation.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation
Claim 23sample processing capabilitysupports2023Source 1needs review

The RT-RPA-LUNAS assay allows SARS-CoV-2 RNA detection without the need for RNA isolation.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation
Claim 24sample processing capabilitysupports2023Source 1needs review

The RT-RPA-LUNAS assay allows SARS-CoV-2 RNA detection without the need for RNA isolation.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation
Claim 25sample processing capabilitysupports2023Source 1needs review

The RT-RPA-LUNAS assay allows SARS-CoV-2 RNA detection without the need for RNA isolation.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation
Claim 26sample processing capabilitysupports2023Source 1needs review

The RT-RPA-LUNAS assay allows SARS-CoV-2 RNA detection without the need for RNA isolation.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation
Claim 27sample processing capabilitysupports2023Source 1needs review

The RT-RPA-LUNAS assay allows SARS-CoV-2 RNA detection without the need for RNA isolation.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation
Claim 28sample processing capabilitysupports2023Source 1needs review

The RT-RPA-LUNAS assay allows SARS-CoV-2 RNA detection without the need for RNA isolation.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation
Claim 29tool descriptionsupports2023Source 1needs review

LUNAS is a bioluminescent nucleic acid sensor platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.

Here, we developed a bioluminescent nucleic acid sensor (LUNAS) platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.
Claim 30tool descriptionsupports2023Source 1needs review

LUNAS is a bioluminescent nucleic acid sensor platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.

Here, we developed a bioluminescent nucleic acid sensor (LUNAS) platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.
Claim 31tool descriptionsupports2023Source 1needs review

LUNAS is a bioluminescent nucleic acid sensor platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.

Here, we developed a bioluminescent nucleic acid sensor (LUNAS) platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.
Claim 32tool descriptionsupports2023Source 1needs review

LUNAS is a bioluminescent nucleic acid sensor platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.

Here, we developed a bioluminescent nucleic acid sensor (LUNAS) platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.
Claim 33tool descriptionsupports2023Source 1needs review

LUNAS is a bioluminescent nucleic acid sensor platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.

Here, we developed a bioluminescent nucleic acid sensor (LUNAS) platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.
Claim 34tool descriptionsupports2023Source 1needs review

LUNAS is a bioluminescent nucleic acid sensor platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.

Here, we developed a bioluminescent nucleic acid sensor (LUNAS) platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.
Claim 35tool descriptionsupports2023Source 1needs review

LUNAS is a bioluminescent nucleic acid sensor platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.

Here, we developed a bioluminescent nucleic acid sensor (LUNAS) platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.
Claim 36use case positioningsupports2023Source 1needs review

RPA-LUNAS is attractive for point-of-care infectious disease testing.

showing that RPA-LUNAS is attractive for point-of-care infectious disease testing.
Claim 37use case positioningsupports2023Source 1needs review

RPA-LUNAS is attractive for point-of-care infectious disease testing.

showing that RPA-LUNAS is attractive for point-of-care infectious disease testing.
Claim 38use case positioningsupports2023Source 1needs review

RPA-LUNAS is attractive for point-of-care infectious disease testing.

showing that RPA-LUNAS is attractive for point-of-care infectious disease testing.
Claim 39use case positioningsupports2023Source 1needs review

RPA-LUNAS is attractive for point-of-care infectious disease testing.

showing that RPA-LUNAS is attractive for point-of-care infectious disease testing.
Claim 40use case positioningsupports2023Source 1needs review

RPA-LUNAS is attractive for point-of-care infectious disease testing.

showing that RPA-LUNAS is attractive for point-of-care infectious disease testing.
Claim 41use case positioningsupports2023Source 1needs review

RPA-LUNAS is attractive for point-of-care infectious disease testing.

showing that RPA-LUNAS is attractive for point-of-care infectious disease testing.
Claim 42use case positioningsupports2023Source 1needs review

RPA-LUNAS is attractive for point-of-care infectious disease testing.

showing that RPA-LUNAS is attractive for point-of-care infectious disease testing.

Approval Evidence

1 source6 linked approval claimsfirst-pass slug lunas
Here, we developed a bioluminescent nucleic acid sensor (LUNAS) platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.

Source:

diagnostic performancesupports

RPA-LUNAS detected SARS-CoV-2 from patient nasopharyngeal swab samples with viral RNA loads of approximately 200 cp/μL within approximately 20 minutes.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation and demonstrated its diagnostic performance for COVID-19 patient nasopharyngeal swab samples. Detection of SARS-CoV-2 from samples with viral RNA loads of ∼200 cp/μL was achieved within ∼20 min

Source:

integration capabilitysupports

LUNAS can be integrated with recombinase polymerase amplification in a rapid one-pot assay with attomolar sensitivity.

LUNAS is easily integrated with recombinase polymerase amplification (RPA), providing attomolar sensitivity in a rapid one-pot assay.

Source:

readout capabilitysupports

A calibrator luciferase enables robust ratiometric readout and real-time monitoring of the RPA reaction using a simple digital camera.

A calibrator luciferase is included for a robust ratiometric readout, enabling real-time monitoring of the RPA reaction using a simple digital camera.

Source:

sample processing capabilitysupports

The RT-RPA-LUNAS assay allows SARS-CoV-2 RNA detection without the need for RNA isolation.

We designed an RT-RPA-LUNAS assay that allows SARS-CoV-2 RNA detection without the need for cumbersome RNA isolation

Source:

tool descriptionsupports

LUNAS is a bioluminescent nucleic acid sensor platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.

Here, we developed a bioluminescent nucleic acid sensor (LUNAS) platform in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes mediating split NanoLuc luciferase complementation.

Source:

use case positioningsupports

RPA-LUNAS is attractive for point-of-care infectious disease testing.

showing that RPA-LUNAS is attractive for point-of-care infectious disease testing.

Source:

Comparisons

Source-backed strengths

The source reports that one-pot RPA-LUNAS achieved attomolar sensitivity and that RPA-LUNAS detected SARS-CoV-2 from patient nasopharyngeal swab samples with viral RNA loads of approximately 200 cp/μL within approximately 20 minutes. A calibrator luciferase enabled robust ratiometric readout and real-time monitoring of the RPA reaction using a simple digital camera.

Ranked Citations

  1. 1.
    StructuralSource 1ACS Central Science2023Claim 1Claim 2Claim 3

    Extracted from this source document.