Toolkit/dCas9-miniRecTE

dCas9-miniRecTE

Multi-Component Switch·Research

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

Summary

we developed a dCas9-miniRecTE editor that enhances large-fragment integration without introducing double-strand breaks in human cells and primary mouse neurons, achieving ∼20% kilobase-scale knock-in efficiency.

Usefulness & Problems

No literature-backed usefulness or problem-fit explainer has been materialized for this record yet.

Published Workflows

Compact bacterial recombination complexes drive efficient kilobase-scale knock-in in mammalian cells.

2026

Objective: Develop compact bacterial recombination-based tools that improve kilobase-scale knock-in and large-fragment integration in mammalian cells.

Why it works: The abstract frames bacterial recombineering proteins as potential solutions to inefficient mammalian HDR and reports that targeted recruitment of EcRecE and engineering of a dCas9-miniRecTE editor improve kilobase-scale integration.

targeted recruitment of bacterial recombineering proteinsfunctional domain engineering of recombination components with dCas9CRISPR/Cas9 targeted recruitmentfunctional domain engineering

Stages

  1. 1.
    EcRecE identification and recruitment-based HDR testing(functional_characterization)

    This stage establishes whether EcRecE is an effective mammalian HDR enhancer before building a more complex editor.

    Selection: Ability of targeted EcRecE recruitment via CRISPR/Cas9 to increase HDR efficiency in mammalian cells.

  2. 2.
    Functional domain engineering of dCas9-miniRecTE(library_design)

    This stage converts the initial HDR-enhancing concept into an engineered editor intended to support DSB-free integration.

    Selection: Combine EcRecE-related engineering with RecT and dCas9 to create a large-fragment integration editor.

  3. 3.
    Cellular validation of dCas9-miniRecTE(confirmatory_validation)

    This stage confirms that the engineered editor functions in human cells and primary mouse neurons for the intended large-fragment integration task.

    Selection: Demonstrate DSB-free kilobase-scale knock-in in relevant mammalian cell contexts.

Steps

  1. 1.
    Test CRISPR/Cas9-targeted recruitment of EcRecE in mammalian HDR assaysHDR-enhancing recombination component

    Determine whether EcRecE can enhance mammalian HDR and kilobase-scale integration.

    The abstract presents EcRecE identification as the first result needed before engineering a more complex editor.

  2. 2.
    Engineer a dCas9-miniRecTE construct from EcRecE-related design, RecT, and dCas9engineered editor

    Create a DSB-free large-fragment integration editor after establishing EcRecE activity.

    The abstract explicitly places editor development after EcRecE identification and ties it to functional domain engineering.

  3. 3.
    Validate dCas9-miniRecTE for DSB-free kilobase-scale knock-in in mammalian cellsvalidated editor

    Confirm that the engineered editor supports large-fragment integration in human cells and primary mouse neurons.

    Validation follows construct development to confirm the intended DSB-free knock-in function in relevant mammalian contexts.

Taxonomy & Function

Primary hierarchy

Mechanism Branch

Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.

Techniques

No technique tags yet.

Target processes

recombination

Validation

Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication

Supporting Sources

Ranked Claims

Claim 1engineering resultsupports2026Source 1needs review

Functional domain engineering of EcRecE with RecT and dCas9 produced dCas9-miniRecTE, a large-fragment integration editor that does not introduce double-strand breaks.

in combination with RecT and a catalytically inactive Cas9 (dCas9), applying functional domain engineering, we developed a dCas9-miniRecTE editor that enhances large-fragment integration without introducing double-strand breaks
Claim 2performancesupports2026Source 1needs review

dCas9-miniRecTE achieved about 20% kilobase-scale knock-in efficiency in human cells and primary mouse neurons.

in human cells and primary mouse neurons, achieving ∼20% kilobase-scale knock-in efficiency
knock-in efficiency 20 %

Approval Evidence

1 source2 linked approval claimsfirst-pass slug dcas9-minirecte
we developed a dCas9-miniRecTE editor that enhances large-fragment integration without introducing double-strand breaks in human cells and primary mouse neurons, achieving ∼20% kilobase-scale knock-in efficiency.

Source:

engineering resultsupports

Functional domain engineering of EcRecE with RecT and dCas9 produced dCas9-miniRecTE, a large-fragment integration editor that does not introduce double-strand breaks.

in combination with RecT and a catalytically inactive Cas9 (dCas9), applying functional domain engineering, we developed a dCas9-miniRecTE editor that enhances large-fragment integration without introducing double-strand breaks

Source:

performancesupports

dCas9-miniRecTE achieved about 20% kilobase-scale knock-in efficiency in human cells and primary mouse neurons.

in human cells and primary mouse neurons, achieving ∼20% kilobase-scale knock-in efficiency

Source:

Comparisons

No literature-backed comparison notes have been materialized for this record yet.

Ranked Citations

  1. 1.
    StructuralSource 1MED2026Claim 1Claim 2

    Extracted from this source document.