Toolkit/AQTrip EL222 variant

AQTrip EL222 variant

Multi-Component Switch·Research·Since 2013

Also known as: AQTrip

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

Summary

AQTrip is an engineered EL222 variant carrying V41I, L52I, A79Q, and V121I substitutions in the blue-light-responsive LOV–HTH transcription factor. It stabilizes the photoactivated state and, in the reported study, oligomerizes without DNA and forms an EL222 dimer–DNA complex in the presence of DNA substrates.

Usefulness & Problems

Why this is useful

AQTrip is useful as a light-responsive protein switch because it prolongs the activated state of EL222 and supports DNA-associated dimer formation under blue-light-responsive control. This makes it relevant for applications that require coupling light input to EL222 DNA-binding behavior and transcription-related regulation.

Problem solved

This variant addresses the problem of limited stability of the EL222 photoactivated state by introducing four point mutations that stabilize that state. The reported behavior also provides a defined DNA-dependent assembly mode, with oligomerization in the absence of DNA and dimer–DNA complex formation when DNA substrates are present.

Problem links

Need conditional recombination or state switching

Derived

AQTrip is an EL222 variant containing V41I, L52I, A79Q, and V121I substitutions that stabilizes the photoactivated state of the blue-light-responsive LOV–HTH protein. In the reported study, this variant oligomerizes in the absence of DNA and forms an EL222 dimer–DNA complex in the presence of DNA substrates.

Need precise spatiotemporal control with light input

Derived

AQTrip is an EL222 variant containing V41I, L52I, A79Q, and V121I substitutions that stabilizes the photoactivated state of the blue-light-responsive LOV–HTH protein. In the reported study, this variant oligomerizes in the absence of DNA and forms an EL222 dimer–DNA complex in the presence of DNA substrates.

Taxonomy & Function

Primary hierarchy

Mechanism Branch

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

Techniques

No technique tags yet.

Target processes

recombination

Input: 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: actuatoroperating role: sensorswitch architecture: multi componentswitch architecture: recruitmentswitch architecture: uncaging

AQTrip is implemented by site-specific substitution of EL222 at V41I, L52I, A79Q, and V121I. The input modality is blue light, and the parent EL222 system operates through reorientation of LOV sensory and HTH effector domains to enable photoactivation of gene transcription.

The available evidence is limited to a single cited study and does not provide quantitative performance metrics in the supplied material. The evidence also does not establish independent replication, detailed cellular validation, or direct demonstration in recombination applications from the provided text.

Validation

Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication

Supporting Sources

Ranked Claims

Claim 1biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 2biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 3biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 4biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 5biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 6biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 7biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 8biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 9biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 10biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 11biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 12biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 13biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 14biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 15biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 16biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 17biophysical behaviorsupports2013Source 1needs review

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
Claim 18engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 19engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 20engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 21engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 22engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 23engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 24engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 25engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 26engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 27engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 28engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 29engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 30engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 31engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 32engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 33engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 34engineering resultsupports2013Source 1needs review

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Claim 35mechanismsupports2013Source 1needs review

EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.

it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
Claim 36mechanismsupports2013Source 1needs review

EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.

it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
Claim 37mechanismsupports2013Source 1needs review

EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.

it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
Claim 38mechanismsupports2013Source 1needs review

EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.

it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
Claim 39mechanismsupports2013Source 1needs review

EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.

it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
Claim 40mechanismsupports2013Source 1needs review

EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.

it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
Claim 41mechanismsupports2013Source 1needs review

EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.

it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
Claim 42mechanismsupports2013Source 1needs review

EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.

it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
Claim 43mechanismsupports2013Source 1needs review

EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.

it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
Claim 44mechanismsupports2013Source 1needs review

EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.

it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
Claim 45mechanistic stepsupports2013Source 1needs review

Blue light induces EL222 dimerization through LOV and HTH interfaces.

Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Claim 46mechanistic stepsupports2013Source 1needs review

Blue light induces EL222 dimerization through LOV and HTH interfaces.

Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Claim 47mechanistic stepsupports2013Source 1needs review

Blue light induces EL222 dimerization through LOV and HTH interfaces.

Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Claim 48mechanistic stepsupports2013Source 1needs review

Blue light induces EL222 dimerization through LOV and HTH interfaces.

Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Claim 49mechanistic stepsupports2013Source 1needs review

Blue light induces EL222 dimerization through LOV and HTH interfaces.

Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Claim 50mechanistic stepsupports2013Source 1needs review

Blue light induces EL222 dimerization through LOV and HTH interfaces.

Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Claim 51mechanistic stepsupports2013Source 1needs review

Blue light induces EL222 dimerization through LOV and HTH interfaces.

Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Claim 52mechanistic stepsupports2013Source 1needs review

Blue light induces EL222 dimerization through LOV and HTH interfaces.

Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Claim 53mechanistic stepsupports2013Source 1needs review

Blue light induces EL222 dimerization through LOV and HTH interfaces.

Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Claim 54mechanistic stepsupports2013Source 1needs review

Blue light induces EL222 dimerization through LOV and HTH interfaces.

Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Claim 55stoichiometrysupports2013Source 1needs review

The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.

NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
Claim 56stoichiometrysupports2013Source 1needs review

The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.

NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
Claim 57stoichiometrysupports2013Source 1needs review

The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.

NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
Claim 58stoichiometrysupports2013Source 1needs review

The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.

NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
Claim 59stoichiometrysupports2013Source 1needs review

The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.

NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
Claim 60stoichiometrysupports2013Source 1needs review

The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.

NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
Claim 61stoichiometrysupports2013Source 1needs review

The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.

NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
Claim 62stoichiometrysupports2013Source 1needs review

The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.

NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
Claim 63stoichiometrysupports2013Source 1needs review

The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.

NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
Claim 64stoichiometrysupports2013Source 1needs review

The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.

NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1

Approval Evidence

1 source2 linked approval claimsfirst-pass slug aqtrip-el222-variant
creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state

Source:

biophysical behaviorsupports

AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.

Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates

Source:

engineering resultsupports

The AQTrip EL222 variant stabilizes the photoactivated state.

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state

Source:

Comparisons

Source-backed strengths

The key reported strength is stabilization of the EL222 photoactivated state in a defined four-mutation variant. The study also directly reports distinct assembly behaviors depending on DNA context, namely oligomerization without DNA and dimer–DNA complex formation with DNA substrates.

Source:

creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state

AQTrip EL222 variant and CRY2-talin/CIBN-CAAX optogenetic plasma membrane recruitment system address a similar problem space because they share recombination.

Shared frame: same top-level item type; shared target processes: recombination; shared mechanisms: heterodimerization; same primary input modality: light

Compared with PA-Cre 3.0

AQTrip EL222 variant and PA-Cre 3.0 address a similar problem space because they share recombination.

Shared frame: same top-level item type; shared target processes: recombination; shared mechanisms: heterodimerization; same primary input modality: light

Compared with PiL[D24]

AQTrip EL222 variant and PiL[D24] address a similar problem space because they share recombination.

Shared frame: same top-level item type; shared target processes: recombination; shared mechanisms: light-induced allosteric switching; same primary input modality: light

Ranked Citations

  1. 1.
    StructuralSource 1Biochemistry2013Claim 1Claim 17Claim 16

    Extracted from this source document.