Source 1primary paper2009Molecular and Cellular Biology
Toolkit/allelic series of Cry mutants
allelic series of Cry mutants
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The allelic series of Cry mutants is a panel of 22 cryptochrome variants, each carrying a single amino acid substitution identified by random mutagenesis and a cell-based screen. The series was used to generate diverse deficiencies across CRY functions and to identify residues involved in protein-protein interaction and CRY2-specific repression.
Usefulness & Problems
Why this is useful
This mutant panel is useful for dissecting cryptochrome structure-function relationships by providing multiple single-residue perturbations with distinct functional defects. It enables comparative analysis of different CRY activities and supports mapping of residues linked to protein-protein interaction and CRY2-specific repression.
Problem solved
This tool addresses the problem of obtaining a diverse set of cryptochrome loss-of-function or separation-of-function variants for mechanistic analysis. The reported mutagenesis and screening strategy produced single-substitution mutants that reveal functionally important residues across different CRY activities.
Problem links
Need better screening or enrichment leverage
DerivedThe allelic series of Cry mutants is a panel of 22 cryptochrome variants, each carrying a single amino acid substitution identified by random mutagenesis and a cell-based screen. The series was used to generate diverse deficiencies across CRY functions and to identify residues involved in protein-protein interaction and CRY2-specific repression.
Need conditional recombination or state switching
DerivedThe allelic series of Cry mutants is a panel of 22 cryptochrome variants, each carrying a single amino acid substitution identified by random mutagenesis and a cell-based screen. The series was used to generate diverse deficiencies across CRY functions and to identify residues involved in protein-protein interaction and CRY2-specific repression.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Mechanisms
altered cry2-specific repressionaltered protein-protein interactionamino acid substitution-based perturbation of protein functionTarget processes
recombinationselectionImplementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: regulator
The series was generated by random mutagenesis followed by a cell-based screen, and each variant contains a single amino acid substitution. The available evidence does not provide construct architecture, expression system details, screening readout design, or delivery considerations.
The provided evidence does not specify the organism, the exact cryptochrome isoform context beyond CRY2-specific repression, or the identities of the substituted residues. It also does not report quantitative performance, validation in multiple assay systems, or independent replication beyond the originating study.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Random mutagenesis followed by a cell-based screen identified 22 Cry mutants with single amino acid substitutions that caused a variety of deficiencies in different CRY functions.
We identified 22 mutants with mutations resulting in single amino acid substitutions which cause a variety of deficiencies in different CRY functions.
identified mutants 22
Random mutagenesis followed by a cell-based screen identified 22 Cry mutants with single amino acid substitutions that caused a variety of deficiencies in different CRY functions.
We identified 22 mutants with mutations resulting in single amino acid substitutions which cause a variety of deficiencies in different CRY functions.
identified mutants 22
Random mutagenesis followed by a cell-based screen identified 22 Cry mutants with single amino acid substitutions that caused a variety of deficiencies in different CRY functions.
We identified 22 mutants with mutations resulting in single amino acid substitutions which cause a variety of deficiencies in different CRY functions.
identified mutants 22
Random mutagenesis followed by a cell-based screen identified 22 Cry mutants with single amino acid substitutions that caused a variety of deficiencies in different CRY functions.
We identified 22 mutants with mutations resulting in single amino acid substitutions which cause a variety of deficiencies in different CRY functions.
identified mutants 22
Random mutagenesis followed by a cell-based screen identified 22 Cry mutants with single amino acid substitutions that caused a variety of deficiencies in different CRY functions.
We identified 22 mutants with mutations resulting in single amino acid substitutions which cause a variety of deficiencies in different CRY functions.
identified mutants 22
Random mutagenesis followed by a cell-based screen identified 22 Cry mutants with single amino acid substitutions that caused a variety of deficiencies in different CRY functions.
We identified 22 mutants with mutations resulting in single amino acid substitutions which cause a variety of deficiencies in different CRY functions.
identified mutants 22
Random mutagenesis followed by a cell-based screen identified 22 Cry mutants with single amino acid substitutions that caused a variety of deficiencies in different CRY functions.
We identified 22 mutants with mutations resulting in single amino acid substitutions which cause a variety of deficiencies in different CRY functions.
identified mutants 22
Approval Evidence
1 source1 linked approval claimfirst-pass slug allelic-series-of-cry-mutants
we created an allelic series of Cry mutants through random mutagenesis
Source:
screening resultsupports
Random mutagenesis followed by a cell-based screen identified 22 Cry mutants with single amino acid substitutions that caused a variety of deficiencies in different CRY functions.
We identified 22 mutants with mutations resulting in single amino acid substitutions which cause a variety of deficiencies in different CRY functions.
Source:
Comparisons
Source-backed strengths
A key strength is that the series comprises 22 independent single amino acid substitution mutants rather than a single defective allele. The source reports that these mutants display a variety of deficiencies in different CRY functions, indicating functional diversity within the panel. The study further links the series to residues involved in protein-protein interaction and CRY2-specific repression.
Compared with cell-free system
allelic series of Cry mutants and cell-free system address a similar problem space because they share recombination, selection.
Shared frame: same top-level item type; shared target processes: recombination, selection
Compared with CfRhPDE1
allelic series of Cry mutants and CfRhPDE1 address a similar problem space because they share recombination, selection.
Shared frame: same top-level item type; shared target processes: recombination, selection
Strengths here: looks easier to implement in practice.
Compared with luciferin-luciferase pair
allelic series of Cry mutants and luciferin-luciferase pair address a similar problem space because they share recombination, selection.
Shared frame: same top-level item type; shared target processes: recombination, selection
Strengths here: looks easier to implement in practice.
Ranked Citations
- 1.