published-canonicalmethodmaha-epistemic/1.0

GUIDE-seq off-target detection

A cell-based sequencing method that captures double-strand oligonucleotides at nuclease-induced DNA breaks to nominate off-target sites. This candidate preserves the experimental unit—editor or circuit, target, cell system, delivery, protocol, assay, comparator, and observation window—before any broader inference is considered.

Bounded definition

A cell-based sequencing method that captures double-strand oligonucleotides at nuclease-induced DNA breaks to nominate off-target sites.

What the cited work establishes

The study develops GUIDE-seq, a cell-based double-strand-break capture method, and applies it to specified nucleases and human cell lines.

The constructs, biological systems, protocols, assays, datasets, and comparisons reported in GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases.

Claims: urn:maha:claim:guide-seq-off-target-detection

What remains a separate question

Assay sensitivity depends on break capture, cell compatibility, sequencing, analysis, and the nuclease modality under study.

GUIDE-seq has assay-specific detection limits and dependencies and does not nominate every possible off-target event in every biological context.

Connected domain graph

Typed dependencies preserve publication state.

Only independently canonical records receive public links and relation statements. Draft graph topology remains private.

mechanistic dependencycanonical

Off-target nomination versus confirmation

outbound connection · comparison

GUIDE-seq produces a nomination set that requires orthogonal confirmation and frequency measurement.

Claim ledger

Every proposition keeps its own evidence state.

empirical-claimsingle-study

The cited study develops GUIDE-seq and applies it to specified RNA-guided nucleases in two human cell lines.

Scope
The constructs, biological systems, protocols, assays, datasets, and comparisons reported in GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases.
Boundary
Assay sensitivity depends on break capture, cell compatibility, sequencing, analysis, and the nuclease modality under study.
Uncertainty
There is no universal effect estimate for this method; numerical results remain attached to the source experiment, biological system, assay, and analysis choices.
Replication
This candidate records one bounded source package. Independent replications and contradictory results must be compiled separately before evidence maturity is upgraded.

Primary sources

Citation, locator, rights, and boundary travel together.

  1. Source 1 · Nature Biotechnology

    GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases

    Shengdar Q. Tsai, Zongli Zheng, Nhu T. Nguyen, Matthew Liebers, et al.

    Exact locator
    Abstract; Figures 1–6; Online Methods; Supplementary Tables; Sequence Read Archive SRP050338.
    Establishes
    The study develops GUIDE-seq, a cell-based double-strand-break capture method, and applies it to specified nucleases and human cell lines.
    Boundary
    GUIDE-seq has assay-specific detection limits and dependencies and does not nominate every possible off-target event in every biological context.
    Rights basis
    citation with paraphrase · Maha paraphrases the source-level result and links to the version of record; no article passage is reproduced.