What question did this study set out to answer?

To develop and validate ICE (Inference of CRISPR Edits), a rapid and low-cost computational assay for analyzing CRISPR editing outcomes using Sanger sequencing data.

January 20, 2018Open Access

Inference of CRISPR Edits from Sanger Trace Data

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Authors

THTim HsiauSynthego (United States)DCDavid S. ConantSynthego (United States)NRNicholas A. RossiSynthego (United States)

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Implication

Computational validation demonstrates reliable CRISPR edit quantification from Sanger trace data, highlighting an accessible alternative to targeted next-generation sequencing.

Key Points

To develop and validate ICE (Inference of CRISPR Edits), a rapid and low-cost computational assay for analyzing CRISPR editing outcomes using Sanger sequencing data.
Modeled potential guide RNA editing outcomes and estimated their presence in Sanger trace data using regression algorithms.
Developed the ICE-Discordance (ICE-D) score to flag unexpected or large structural edits.
Validated the software empirically across >1,800 editing experiments and benchmarked results against next-generation amplicon sequencing (Amp-Seq).
Demonstrated robust and reproducible analysis across >1,800 edits, with estimates strongly correlating with Amp-Seq data.
Successfully identified complex editing events, including multi-guide cuts and homology-directed repair outcomes such as base edits and knock-ins.

Cite This Study

Hsiau et al. (2018) studied this question.

synapsesocial.com/papers/6a7660e3232067cbf2ec0040 https://doi.org/10.1101/251082

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Implication

Computational validation demonstrates reliable CRISPR edit quantification from Sanger trace data, highlighting an accessible alternative to targeted next-generation sequencing.

Inference of CRISPR Edits from Sanger Trace Data

Discussion

Member takes

Implication

Key Points

Cite This Study

Inference of CRISPR Edits from Sanger Trace Data

Discussion

Member takes

Implication

Key Points

Cite This Study

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