Toolkit/photo-crosslinking

photo-crosslinking

Construct Pattern·Research·Since 2021

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

Summary

Photo-crosslinking in this context is an application of genetic code expansion in Bacillus subtilis that enables light-triggered covalent capture of molecular interactions. The reported system was part of a broader noncanonical amino acid incorporation platform used for photo-crosslinking, click-labelling, and translational titration.

Usefulness & Problems

Why this is useful

This approach is useful for probing protein interactions and cell-division biology in vivo in Bacillus subtilis using genetically encoded, light-responsive chemistry. The source study indicates that the underlying genetic code expansion platform enabled interrogation of bacterial cytokinesis by precise modulation of cell division dynamics.

Source:

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.

Source:

we demonstrate broad and efficient genetic code expansion in B. subtilis by incorporating 20 distinct non-standard amino acids within proteins using 3 different families of genetic code expansion systems and two choices of codons

Problem solved

It addresses the problem of introducing photo-reactive chemical functionality into proteins in Bacillus subtilis, where efficient genetic code expansion had been limited. This enables covalent trapping of interactions under light control within a bacterial system.

Source:

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.

Problem links

Need precise spatiotemporal control with light input

Derived

Photo-crosslinking is an application enabled by genetic code expansion systems in Bacillus subtilis, where light-responsive noncanonical amino acid incorporation is used to support covalent capture of molecular interactions. In the cited study, these systems were used alongside click-labeling and translational titration and were applied to interrogate bacterial cell division and validate a predicted protein-protein binding interface.

Need tighter control over protein production

Derived

Photo-crosslinking is an application enabled by genetic code expansion systems in Bacillus subtilis, where light-responsive noncanonical amino acid incorporation is used to support covalent capture of molecular interactions. In the cited study, these systems were used alongside click-labeling and translational titration and were applied to interrogate bacterial cell division and validate a predicted protein-protein binding interface.

Taxonomy & Function

Primary hierarchy

Mechanism Branch

Architecture: A reusable architecture pattern for arranging parts into an engineered system.

Techniques

No technique tags yet.

Target processes

translation

Input: Light

Implementation Constraints

cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: spectral hardware requirementoperating role: builder

Implementation depends on a genetic code expansion system in Bacillus subtilis capable of incorporating noncanonical amino acids, and the study reports use of three system families and two codon choices. The supplied evidence does not provide construct architecture, orthogonal synthetase/tRNA identities, or optical exposure parameters for the photo-crosslinking application.

The provided evidence does not specify the photo-crosslinking amino acid, illumination wavelength, crosslinking efficiency, or target proteins. Validation described here is limited to a single 2021 study in Bacillus subtilis, with no independent replication provided.

Validation

Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication

Observations

successBacteriaapplication demoBacillus subtilis

Inferred from claim c2 during normalization. The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration. Derived from claim c2. Quoted text: We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.

Source:

successBacteriaapplication demoBacillus subtilis

Inferred from claim c2 during normalization. The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration. Derived from claim c2. Quoted text: We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.

Source:

successBacteriaapplication demoBacillus subtilis

Inferred from claim c2 during normalization. The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration. Derived from claim c2. Quoted text: We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.

Source:

successBacteriaapplication demoBacillus subtilis

Inferred from claim c2 during normalization. The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration. Derived from claim c2. Quoted text: We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.

Source:

successBacteriaapplication demoBacillus subtilis

Inferred from claim c2 during normalization. The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration. Derived from claim c2. Quoted text: We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.

Source:

successBacteriaapplication demoBacillus subtilis

Inferred from claim c2 during normalization. The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration. Derived from claim c2. Quoted text: We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.

Source:

successBacteriaapplication demoBacillus subtilis

Inferred from claim c2 during normalization. The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration. Derived from claim c2. Quoted text: We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.

Source:

Supporting Sources

Ranked Claims

Claim 1applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 2applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 3applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 4applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 5applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 6applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 7applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 8applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 9applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 10applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 11applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 12applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 13applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 14applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 15applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 16applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 17applicationsupports2021Source 1needs review

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.
Claim 18biological applicationsupports2021Source 1needs review

These tools were used to begin interrogating properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo.

begin to interrogate properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo
Claim 19biological applicationsupports2021Source 1needs review

These tools were used to begin interrogating properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo.

begin to interrogate properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo
Claim 20biological applicationsupports2021Source 1needs review

These tools were used to begin interrogating properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo.

begin to interrogate properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo
Claim 21biological applicationsupports2021Source 1needs review

These tools were used to begin interrogating properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo.

begin to interrogate properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo
Claim 22biological applicationsupports2021Source 1needs review

These tools were used to begin interrogating properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo.

begin to interrogate properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo
Claim 23biological applicationsupports2021Source 1needs review

These tools were used to begin interrogating properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo.

begin to interrogate properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo
Claim 24biological applicationsupports2021Source 1needs review

These tools were used to begin interrogating properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo.

begin to interrogate properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo
Claim 25biological applicationsupports2021Source 1needs review

These tools were used to begin interrogating properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo.

begin to interrogate properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo
Claim 26biological applicationsupports2021Source 1needs review

These tools were used to begin interrogating properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo.

begin to interrogate properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo
Claim 27biological applicationsupports2021Source 1needs review

These tools were used to begin interrogating properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo.

begin to interrogate properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo
Claim 28capabilitysupports2021Source 1needs review

The authors demonstrate broad and efficient genetic code expansion in Bacillus subtilis using 3 families of genetic code expansion systems and 2 codon choices.

we demonstrate broad and efficient genetic code expansion in B. subtilis by incorporating 20 distinct non-standard amino acids within proteins using 3 different families of genetic code expansion systems and two choices of codons
codon choices 2distinct non-standard amino acids incorporated 20genetic code expansion system families 3
Claim 29capabilitysupports2021Source 1needs review

The authors demonstrate broad and efficient genetic code expansion in Bacillus subtilis using 3 families of genetic code expansion systems and 2 codon choices.

we demonstrate broad and efficient genetic code expansion in B. subtilis by incorporating 20 distinct non-standard amino acids within proteins using 3 different families of genetic code expansion systems and two choices of codons
codon choices 2distinct non-standard amino acids incorporated 20genetic code expansion system families 3
Claim 30capabilitysupports2021Source 1needs review

The authors demonstrate broad and efficient genetic code expansion in Bacillus subtilis using 3 families of genetic code expansion systems and 2 codon choices.

we demonstrate broad and efficient genetic code expansion in B. subtilis by incorporating 20 distinct non-standard amino acids within proteins using 3 different families of genetic code expansion systems and two choices of codons
codon choices 2distinct non-standard amino acids incorporated 20genetic code expansion system families 3
Claim 31capabilitysupports2021Source 1needs review

The authors demonstrate broad and efficient genetic code expansion in Bacillus subtilis using 3 families of genetic code expansion systems and 2 codon choices.

we demonstrate broad and efficient genetic code expansion in B. subtilis by incorporating 20 distinct non-standard amino acids within proteins using 3 different families of genetic code expansion systems and two choices of codons
codon choices 2distinct non-standard amino acids incorporated 20genetic code expansion system families 3
Claim 32capabilitysupports2021Source 1needs review

The authors demonstrate broad and efficient genetic code expansion in Bacillus subtilis using 3 families of genetic code expansion systems and 2 codon choices.

we demonstrate broad and efficient genetic code expansion in B. subtilis by incorporating 20 distinct non-standard amino acids within proteins using 3 different families of genetic code expansion systems and two choices of codons
codon choices 2distinct non-standard amino acids incorporated 20genetic code expansion system families 3
Claim 33comparative observationsupports2021Source 1needs review

These tools allowed the authors to demonstrate differences between E. coli and Bacillus subtilis stop codon suppression.

These tools allow us to demonstrate differences between E. coli and B. subtilis stop codon suppression
Claim 34comparative observationsupports2021Source 1needs review

These tools allowed the authors to demonstrate differences between E. coli and Bacillus subtilis stop codon suppression.

These tools allow us to demonstrate differences between E. coli and B. subtilis stop codon suppression
Claim 35comparative observationsupports2021Source 1needs review

These tools allowed the authors to demonstrate differences between E. coli and Bacillus subtilis stop codon suppression.

These tools allow us to demonstrate differences between E. coli and B. subtilis stop codon suppression
Claim 36comparative observationsupports2021Source 1needs review

These tools allowed the authors to demonstrate differences between E. coli and Bacillus subtilis stop codon suppression.

These tools allow us to demonstrate differences between E. coli and B. subtilis stop codon suppression
Claim 37comparative observationsupports2021Source 1needs review

These tools allowed the authors to demonstrate differences between E. coli and Bacillus subtilis stop codon suppression.

These tools allow us to demonstrate differences between E. coli and B. subtilis stop codon suppression
Claim 38study focussupports2021Source 1needs review

The paper concerns designing efficient genetic code expansion in Bacillus subtilis to gain biological insights.

Designing efficient genetic code expansion in Bacillus subtilis to gain biological insights
Section: title
Claim 39study focussupports2021Source 1needs review

The paper concerns designing efficient genetic code expansion in Bacillus subtilis to gain biological insights.

Designing efficient genetic code expansion in Bacillus subtilis to gain biological insights
Section: title
Claim 40study focussupports2021Source 1needs review

The paper concerns designing efficient genetic code expansion in Bacillus subtilis to gain biological insights.

Designing efficient genetic code expansion in Bacillus subtilis to gain biological insights
Section: title
Claim 41study focussupports2021Source 1needs review

The paper concerns designing efficient genetic code expansion in Bacillus subtilis to gain biological insights.

Designing efficient genetic code expansion in Bacillus subtilis to gain biological insights
Section: title
Claim 42study focussupports2021Source 1needs review

The paper concerns designing efficient genetic code expansion in Bacillus subtilis to gain biological insights.

Designing efficient genetic code expansion in Bacillus subtilis to gain biological insights
Section: title
Claim 43validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 44validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 45validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 46validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 47validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 48validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 49validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 50validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 51validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 52validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 53validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 54validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 55validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 56validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 57validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 58validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface
Claim 59validation usesupports2021Source 1needs review

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface

Approval Evidence

1 source2 linked approval claimsfirst-pass slug photo-crosslinking
We use these systems to achieve ... photo-crosslinking

Source:

applicationsupports

The genetic code expansion systems were used to achieve click-labelling, photo-crosslinking, and translational titration.

We use these systems to achieve click-labelling, photo-crosslinking, and translational titration.

Source:

validation usesupports

These tools were used to validate a predicted protein-protein binding interface.

validate a predicted protein-protein binding interface

Source:

Comparisons

Source-backed strengths

The source study reports broad and efficient genetic code expansion in Bacillus subtilis using three families of systems and two codon choices, providing the foundation for photo-crosslinking applications. Photo-crosslinking was demonstrated as one of several enabled downstream uses of this platform.

Source:

These tools allow us to demonstrate differences between E. coli and B. subtilis stop codon suppression

Compared with click-labelling

photo-crosslinking and click-labelling address a similar problem space because they share translation.

Shared frame: same top-level item type; shared target processes: translation; shared mechanisms: genetic code expansion, translation control, translation_control; same primary input modality: light

photo-crosslinking and genetic code expansion in Bacillus subtilis address a similar problem space because they share translation.

Shared frame: same top-level item type; shared target processes: translation; shared mechanisms: translation control, translation_control; same primary input modality: light

Relative tradeoffs: looks easier to implement in practice.

photo-crosslinking and translational titration address a similar problem space because they share translation.

Shared frame: same top-level item type; shared target processes: translation; shared mechanisms: genetic code expansion, translation control, translation_control; same primary input modality: light

Relative tradeoffs: looks easier to implement in practice.

Ranked Citations

  1. 1.
    StructuralSource 1Nature Communications2021Claim 1Claim 16Claim 17

    Extracted from this source document.