Source 1primary paper2018
Toolkit/split APEX peroxidase
split APEX peroxidase
Also known as: sAPEX
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Split APEX peroxidase (sAPEX) is an engineered two-fragment version of APEX generated by directed evolution. It consists of a 200-amino acid N-terminal fragment (AP) and a 50-amino acid C-terminal fragment (EX) that can reconstitute peroxidase activity in mammalian cells.
Usefulness & Problems
Why this is useful
sAPEX enables conditional APEX peroxidase activity only where its two fragments are brought together. The reported demonstrations in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites indicate utility for spatially restricted labeling or detection at defined intracellular locales.
Source:
We demonstrate sAPEX reconstitution in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
Problem solved
sAPEX addresses the problem of restricting APEX activity to specific molecular or subcellular contexts that are difficult to target with a single intact enzyme. The split design allows activity to be reconstituted at selected sites such as engineered RNA motifs and organelle contact sites.
Source:
We demonstrate sAPEX reconstitution in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
split-enzyme reconstitutionTechniques
Directed EvolutionTarget processes
No target processes tagged yet.
Implementation Constraints
The evolved system comprises two fragments: AP, a 200-amino acid N-terminal fragment, and EX, a 50-amino acid C-terminal fragment. The supplied evidence supports implementation in mammalian cells, but it does not provide construct architecture, expression conditions, or cofactor requirements.
The supplied evidence does not report quantitative performance metrics, background activity, kinetics, or comparison to full-length APEX. Independent replication is not provided in the supplied evidence, and validation is limited to the contexts described in the source study.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
sAPEX reconstitution was demonstrated in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
We demonstrate sAPEX reconstitution in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
sAPEX reconstitution was demonstrated in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
We demonstrate sAPEX reconstitution in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
sAPEX reconstitution was demonstrated in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
We demonstrate sAPEX reconstitution in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
sAPEX reconstitution was demonstrated in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
We demonstrate sAPEX reconstitution in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
sAPEX reconstitution was demonstrated in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
We demonstrate sAPEX reconstitution in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
sAPEX reconstitution was demonstrated in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
We demonstrate sAPEX reconstitution in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
sAPEX reconstitution was demonstrated in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
We demonstrate sAPEX reconstitution in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
The evolved split APEX system comprises a 200-amino acid N-terminal fragment called AP and a 50-amino acid C-terminal fragment called EX.
produced a 200-amino acid N-terminal fragment (with 9 mutations relative to APEX2) called “AP” and a 50-amino acid C-terminal fragment called “EX”
The evolved split APEX system comprises a 200-amino acid N-terminal fragment called AP and a 50-amino acid C-terminal fragment called EX.
produced a 200-amino acid N-terminal fragment (with 9 mutations relative to APEX2) called “AP” and a 50-amino acid C-terminal fragment called “EX”
The evolved split APEX system comprises a 200-amino acid N-terminal fragment called AP and a 50-amino acid C-terminal fragment called EX.
produced a 200-amino acid N-terminal fragment (with 9 mutations relative to APEX2) called “AP” and a 50-amino acid C-terminal fragment called “EX”
The evolved split APEX system comprises a 200-amino acid N-terminal fragment called AP and a 50-amino acid C-terminal fragment called EX.
produced a 200-amino acid N-terminal fragment (with 9 mutations relative to APEX2) called “AP” and a 50-amino acid C-terminal fragment called “EX”
The evolved split APEX system comprises a 200-amino acid N-terminal fragment called AP and a 50-amino acid C-terminal fragment called EX.
produced a 200-amino acid N-terminal fragment (with 9 mutations relative to APEX2) called “AP” and a 50-amino acid C-terminal fragment called “EX”
The evolved split APEX system comprises a 200-amino acid N-terminal fragment called AP and a 50-amino acid C-terminal fragment called EX.
produced a 200-amino acid N-terminal fragment (with 9 mutations relative to APEX2) called “AP” and a 50-amino acid C-terminal fragment called “EX”
The evolved split APEX system comprises a 200-amino acid N-terminal fragment called AP and a 50-amino acid C-terminal fragment called EX.
produced a 200-amino acid N-terminal fragment (with 9 mutations relative to APEX2) called “AP” and a 50-amino acid C-terminal fragment called “EX”
Directed evolution produced a split APEX tool named sAPEX.
we used directed evolution to engineer a split APEX tool (sAPEX)
Directed evolution produced a split APEX tool named sAPEX.
we used directed evolution to engineer a split APEX tool (sAPEX)
Directed evolution produced a split APEX tool named sAPEX.
we used directed evolution to engineer a split APEX tool (sAPEX)
Directed evolution produced a split APEX tool named sAPEX.
we used directed evolution to engineer a split APEX tool (sAPEX)
Directed evolution produced a split APEX tool named sAPEX.
we used directed evolution to engineer a split APEX tool (sAPEX)
Directed evolution produced a split APEX tool named sAPEX.
we used directed evolution to engineer a split APEX tool (sAPEX)
Directed evolution produced a split APEX tool named sAPEX.
we used directed evolution to engineer a split APEX tool (sAPEX)
Approval Evidence
1 source3 linked approval claimsfirst-pass slug split-apex-peroxidase
we used directed evolution to engineer a split APEX tool (sAPEX)
Source:
application demosupports
sAPEX reconstitution was demonstrated in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
We demonstrate sAPEX reconstitution in the mammalian cytosol, on engineered RNA motifs within telomerase noncoding RNA, and at mitochondria-endoplasmic reticulum contact sites.
Source:
compositionsupports
The evolved split APEX system comprises a 200-amino acid N-terminal fragment called AP and a 50-amino acid C-terminal fragment called EX.
produced a 200-amino acid N-terminal fragment (with 9 mutations relative to APEX2) called “AP” and a 50-amino acid C-terminal fragment called “EX”
Source:
engineering resultsupports
Directed evolution produced a split APEX tool named sAPEX.
we used directed evolution to engineer a split APEX tool (sAPEX)
Source:
Comparisons
Source-backed strengths
The tool was produced by directed evolution and was reported to reconstitute activity in mammalian cells. Its use was demonstrated across multiple intracellular contexts, including the cytosol, telomerase noncoding RNA bearing engineered motifs, and mitochondria-endoplasmic reticulum contact sites.
Source:
we used directed evolution to engineer a split APEX tool (sAPEX)
Ranked Citations
- 1.