Source 1primary paper2023
Toolkit/light-inducible split Cre recombinase
light-inducible split Cre recombinase
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The light-inducible split Cre recombinase is an optogenetic multi-component switch in which split Cre recombinase fragments are coupled to light-inducible dimerization modules to achieve inducible post-translational control of Cre activity. It was characterized by comprehensive screening of split sites across the Cre protein using a pooled, sequencing-based domain insertion profiling approach.
Usefulness & Problems
Why this is useful
This tool is useful for identifying and implementing light-gated split Cre configurations that enable temporal control of site-specific recombination at the protein level. The associated screening framework also provides a streamlined route to engineer inducible post-translational control for proteins of interest.
Source:
We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on split sites throughout the protein.
Source:
To improve accuracy in predicting split protein behavior, we develop a Bayesian computational approach to contextualize errors inherent to experimental procedures.
Source:
we use a pooled library approach that enables rapid generation and screening of nearly all possible split protein constructs in parallel, where results can be read out using sequencing
Problem solved
It addresses the problem of finding functional split sites in Cre recombinase that can be converted into a light-inducible switch rather than relying on sparse, manually chosen split configurations. It also addresses the experimental bottleneck of evaluating large numbers of split-protein designs by enabling parallel pooled screening with sequencing-based readout.
Source:
We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on split sites throughout the protein.
Problem links
Need better screening or enrichment leverage
DerivedThe light-inducible split Cre recombinase is a multi-component optogenetic switch in which Cre recombinase was evaluated in split configurations coupled to optogenetic dimerization. It enables light-inducible post-translational control of Cre-mediated recombination and was characterized through comprehensive mapping of split sites across the protein.
Need conditional recombination or state switching
DerivedThe light-inducible split Cre recombinase is a multi-component optogenetic switch in which Cre recombinase was evaluated in split configurations coupled to optogenetic dimerization. It enables light-inducible post-translational control of Cre-mediated recombination and was characterized through comprehensive mapping of split sites across the protein.
Need precise spatiotemporal control with light input
DerivedThe light-inducible split Cre recombinase is a multi-component optogenetic switch in which Cre recombinase was evaluated in split configurations coupled to optogenetic dimerization. It enables light-inducible post-translational control of Cre-mediated recombination and was characterized through comprehensive mapping of split sites across the protein.
Need tighter control over protein production
DerivedThe light-inducible split Cre recombinase is a multi-component optogenetic switch in which Cre recombinase was evaluated in split configurations coupled to optogenetic dimerization. It enables light-inducible post-translational control of Cre-mediated recombination and was characterized through comprehensive mapping of split sites across the protein.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
light-induced dimerizationlight-induced dimerizationpost-translational controlpost-translational controlsite-specific recombinationsite-specific recombinationsplit-protein reconstitutionsplit-protein reconstitutionTranslation ControlTarget processes
recombinationselectiontranslationInput: 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: regulatoroperating role: sensorswitch architecture: multi componentswitch architecture: split
Implementation involves expressing split Cre recombinase fragments fused to optogenetic dimerization partners so that light can induce reconstitution-dependent Cre function. The reported engineering and evaluation relied on a pooled library, sequencing-based readout, and Bayesian computational modeling, but the supplied evidence does not specify construct architecture, host system, or optical parameters.
The available evidence is limited to a single 2023 source focused on screening and mapping split-site behavior in Cre recombinase. Specific optogenetic dimer pairs, illumination wavelengths, recombination performance metrics, and validation across organisms or application settings are not provided in the supplied evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Applying the method to Cre recombinase with optogenetic dimers produced comprehensive data on split sites throughout the protein.
We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on split sites throughout the protein.
Applying the method to Cre recombinase with optogenetic dimers produced comprehensive data on split sites throughout the protein.
We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on split sites throughout the protein.
Applying the method to Cre recombinase with optogenetic dimers produced comprehensive data on split sites throughout the protein.
We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on split sites throughout the protein.
Applying the method to Cre recombinase with optogenetic dimers produced comprehensive data on split sites throughout the protein.
We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on split sites throughout the protein.
Applying the method to Cre recombinase with optogenetic dimers produced comprehensive data on split sites throughout the protein.
We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on split sites throughout the protein.
Applying the method to Cre recombinase with optogenetic dimers produced comprehensive data on split sites throughout the protein.
We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on split sites throughout the protein.
Applying the method to Cre recombinase with optogenetic dimers produced comprehensive data on split sites throughout the protein.
We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on split sites throughout the protein.
Applying the method to Cre recombinase with optogenetic dimers produced comprehensive data on split sites throughout the protein.
We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on split sites throughout the protein.
Applying the method to Cre recombinase with optogenetic dimers produced comprehensive data on split sites throughout the protein.
We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on split sites throughout the protein.
Applying the method to Cre recombinase with optogenetic dimers produced comprehensive data on split sites throughout the protein.
We perform our method on Cre recombinase with optogenetic dimers as a proof of concept, resulting in comprehensive data on split sites throughout the protein.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
A Bayesian computational approach was developed to improve accuracy in predicting split protein behavior by contextualizing errors inherent to experimental procedures.
To improve accuracy in predicting split protein behavior, we develop a Bayesian computational approach to contextualize errors inherent to experimental procedures.
A Bayesian computational approach was developed to improve accuracy in predicting split protein behavior by contextualizing errors inherent to experimental procedures.
To improve accuracy in predicting split protein behavior, we develop a Bayesian computational approach to contextualize errors inherent to experimental procedures.
A Bayesian computational approach was developed to improve accuracy in predicting split protein behavior by contextualizing errors inherent to experimental procedures.
To improve accuracy in predicting split protein behavior, we develop a Bayesian computational approach to contextualize errors inherent to experimental procedures.
A Bayesian computational approach was developed to improve accuracy in predicting split protein behavior by contextualizing errors inherent to experimental procedures.
To improve accuracy in predicting split protein behavior, we develop a Bayesian computational approach to contextualize errors inherent to experimental procedures.
A Bayesian computational approach was developed to improve accuracy in predicting split protein behavior by contextualizing errors inherent to experimental procedures.
To improve accuracy in predicting split protein behavior, we develop a Bayesian computational approach to contextualize errors inherent to experimental procedures.
A Bayesian computational approach was developed to improve accuracy in predicting split protein behavior by contextualizing errors inherent to experimental procedures.
To improve accuracy in predicting split protein behavior, we develop a Bayesian computational approach to contextualize errors inherent to experimental procedures.
A Bayesian computational approach was developed to improve accuracy in predicting split protein behavior by contextualizing errors inherent to experimental procedures.
To improve accuracy in predicting split protein behavior, we develop a Bayesian computational approach to contextualize errors inherent to experimental procedures.
A Bayesian computational approach was developed to improve accuracy in predicting split protein behavior by contextualizing errors inherent to experimental procedures.
To improve accuracy in predicting split protein behavior, we develop a Bayesian computational approach to contextualize errors inherent to experimental procedures.
A Bayesian computational approach was developed to improve accuracy in predicting split protein behavior by contextualizing errors inherent to experimental procedures.
To improve accuracy in predicting split protein behavior, we develop a Bayesian computational approach to contextualize errors inherent to experimental procedures.
A Bayesian computational approach was developed to improve accuracy in predicting split protein behavior by contextualizing errors inherent to experimental procedures.
To improve accuracy in predicting split protein behavior, we develop a Bayesian computational approach to contextualize errors inherent to experimental procedures.
A pooled library approach enables rapid generation and screening of nearly all possible split protein constructs in parallel, with sequencing-based readout.
we use a pooled library approach that enables rapid generation and screening of nearly all possible split protein constructs in parallel, where results can be read out using sequencing
A pooled library approach enables rapid generation and screening of nearly all possible split protein constructs in parallel, with sequencing-based readout.
we use a pooled library approach that enables rapid generation and screening of nearly all possible split protein constructs in parallel, where results can be read out using sequencing
A pooled library approach enables rapid generation and screening of nearly all possible split protein constructs in parallel, with sequencing-based readout.
we use a pooled library approach that enables rapid generation and screening of nearly all possible split protein constructs in parallel, where results can be read out using sequencing
A pooled library approach enables rapid generation and screening of nearly all possible split protein constructs in parallel, with sequencing-based readout.
we use a pooled library approach that enables rapid generation and screening of nearly all possible split protein constructs in parallel, where results can be read out using sequencing
A pooled library approach enables rapid generation and screening of nearly all possible split protein constructs in parallel, with sequencing-based readout.
we use a pooled library approach that enables rapid generation and screening of nearly all possible split protein constructs in parallel, where results can be read out using sequencing
A pooled library approach enables rapid generation and screening of nearly all possible split protein constructs in parallel, with sequencing-based readout.
we use a pooled library approach that enables rapid generation and screening of nearly all possible split protein constructs in parallel, where results can be read out using sequencing
A pooled library approach enables rapid generation and screening of nearly all possible split protein constructs in parallel, with sequencing-based readout.
we use a pooled library approach that enables rapid generation and screening of nearly all possible split protein constructs in parallel, where results can be read out using sequencing
A pooled library approach enables rapid generation and screening of nearly all possible split protein constructs in parallel, with sequencing-based readout.
we use a pooled library approach that enables rapid generation and screening of nearly all possible split protein constructs in parallel, where results can be read out using sequencing
A pooled library approach enables rapid generation and screening of nearly all possible split protein constructs in parallel, with sequencing-based readout.
we use a pooled library approach that enables rapid generation and screening of nearly all possible split protein constructs in parallel, where results can be read out using sequencing
A pooled library approach enables rapid generation and screening of nearly all possible split protein constructs in parallel, with sequencing-based readout.
we use a pooled library approach that enables rapid generation and screening of nearly all possible split protein constructs in parallel, where results can be read out using sequencing
Approval Evidence
1 source1 linked approval claimfirst-pass slug light-inducible-split-cre-recombinase
Comprehensive screening of a light-inducible split Cre recombinase with domain insertion profiling
Source:
method advantagesupports
The overall method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Overall, our method provides a streamlined approach for achieving inducible post-translational control of a protein of interest.
Source:
Comparisons
Source-backed strengths
The reported work produced comprehensive data on split sites throughout Cre recombinase when combined with optogenetic dimers. The method supports rapid generation and screening of nearly all possible split constructs in parallel, and a Bayesian computational approach was developed to improve prediction accuracy by accounting for experimental error.
Compared with cLIPS2
light-inducible split Cre recombinase and cLIPS2 address a similar problem space because they share recombination, selection, translation.
Shared frame: same top-level item type; shared target processes: recombination, selection, translation; shared mechanisms: translation_control; same primary input modality: light
Compared with CRISPR/Cas9 system
light-inducible split Cre recombinase and CRISPR/Cas9 system address a similar problem space because they share recombination, selection, translation.
Shared frame: same top-level item type; shared target processes: recombination, selection, translation; shared mechanisms: translation_control
Relative tradeoffs: appears more independently replicated; looks easier to implement in practice.
Compared with pooled library approach
light-inducible split Cre recombinase and pooled library approach address a similar problem space because they share recombination, selection, translation.
Shared frame: shared target processes: recombination, selection, translation; shared mechanisms: translation_control; same primary input modality: light
Relative tradeoffs: looks easier to implement in practice.
Ranked Citations
- 1.