Source 1primary paper2023
Toolkit/pooled library approach
pooled library approach
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The pooled library approach is an engineering method for rapid generation and parallel screening of nearly all possible split-protein constructs, with sequencing-based readout. In the cited application, it was used with optogenetic dimers to comprehensively map split-site behavior across Cre recombinase and support inducible post-translational control design.
Usefulness & Problems
Why this is useful
This approach is useful for systematically exploring split-protein design space at high coverage rather than testing a small number of manually chosen constructs. The cited study indicates that it streamlines development of inducible post-translational control systems 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 difficulty of identifying functional split sites across a protein when the number of possible constructs is large. The method enables parallel generation and screening of nearly all possible split variants with sequencing-based readout, reducing the need for one-by-one construct testing.
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 pooled library approach is an engineering method for rapid generation and parallel screening of nearly all possible split-protein constructs, with sequencing-based readout. In the cited application, it was used with optogenetic dimers to comprehensively map split-site behavior across Cre recombinase and support inducible post-translational control design.
Need conditional recombination or state switching
DerivedThe pooled library approach is an engineering method for rapid generation and parallel screening of nearly all possible split-protein constructs, with sequencing-based readout. In the cited application, it was used with optogenetic dimers to comprehensively map split-site behavior across Cre recombinase and support inducible post-translational control design.
Need precise spatiotemporal control with light input
DerivedThe pooled library approach is an engineering method for rapid generation and parallel screening of nearly all possible split-protein constructs, with sequencing-based readout. In the cited application, it was used with optogenetic dimers to comprehensively map split-site behavior across Cre recombinase and support inducible post-translational control design.
Need tighter control over protein production
DerivedThe pooled library approach is an engineering method for rapid generation and parallel screening of nearly all possible split-protein constructs, with sequencing-based readout. In the cited application, it was used with optogenetic dimers to comprehensively map split-site behavior across Cre recombinase and support inducible post-translational control design.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete method used to build, optimize, or evolve an engineered system.
Target 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: builderswitch architecture: multi componentswitch architecture: recruitmentswitch architecture: split
The reported implementation used sequencing as the assay readout for pooled screening of split-protein constructs. In the cited application, the method was paired with optogenetic dimers and applied to Cre recombinase, but the provided evidence does not detail construct architecture, host system, or illumination parameters.
The supplied evidence is limited to a single 2023 source and one application in Cre recombinase. The evidence does not specify quantitative performance metrics, generality across other proteins, or operational details of library construction and screening.
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.
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 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 source3 linked approval claimsfirst-pass slug pooled-library-approach
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
Source:
application resultsupports
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.
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:
method capabilitysupports
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
Source:
Comparisons
Source-backed strengths
A key strength is the ability to generate and screen nearly all possible split-protein constructs in parallel. In the reported application, it produced comprehensive data on split sites throughout Cre recombinase when combined with optogenetic dimers.
Compared with 3D bioprinting
pooled library approach and 3D bioprinting 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 cLIPS2
pooled library approach and cLIPS2 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
Strengths here: looks easier to implement in practice.
Compared with light-inducible split Cre recombinase
pooled library approach and light-inducible split Cre recombinase 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
Strengths here: looks easier to implement in practice.
Ranked Citations
- 1.