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Experimental biology · CRISPR-Cas9 gene-editing study

CRISPR-Cas9 gene-editing study

How do you know a targeted edit actually happened?

Project account · methods and observations from my study and research notes.

01 / The question

How do you know a targeted edit actually happened?

At the LADE laboratory of Universidad de Concepción, the May–August 2025 internship focused on targeted mutagenesis in Xenopus tropicalis. The engineering challenge was to connect a bioinformatic target to an interpretable experimental result.

Sequence → target → validation
Conceptual illustration of the approach

02 / The intuition

A mechanism worth testing.

A visible phenotype and a sequencing result answer different questions. The workflow connected guide design, experiment tracking and genotyping rather than treating any one readout as sufficient.

03 / The work

Inside the method.

Explore each part of the approach.

01Inspect the target

Comparative sequence analysis and guide-design tools supported target selection. The profile records work with BLAST, Clustal, CHOPCHOP, Benchling and CRISPRscan.

02Connect design to experiment

The internship covered guide preparation and experimental delivery, with attention to documenting a repeatable workflow and interpreting mosaic outcomes.

03Check the edit

Sanger sequencing and ICE analysis were used alongside phenotype observations to assess the resulting edits.

04 / The observations

What emerged.

The profile records mosaic depigmentation phenotypes and Sanger/ICE-based genotyping. These are complementary observations: one describes an organism-level outcome, the other the sequence mixture in the analysed sample.

This account is based on recorded project notes. Original reports, figures and datasets are not embedded here.

05 / The limits

Where the evidence stops.

The original sequencing traces, sample counts and guide-specific results are needed to substantiate an editing-efficiency figure. Mosaicism and genotype–phenotype interpretation should remain explicit.

The academic foundations

Where this work connects.

Biology & biotechnologyProgramming & computational methods
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