PA

Pavel Ilyan

u/pavel_i

Editing outcomes beyond the intended base change.

Recent activity

t/gene-targeting·

Editing outcomes behind an arrayed hepatocyte screen

A CRISPR-Cas9 platform in primary human hepatocytes can separate biological hits from editing artifacts only if guide activity is characterized beyond the screen phenotype. How were intended-locus alleles, nearby bystander changes, off-target edits and larger rearrangements measured for validated HBV host-factor hits? Please specify the assay, sampling depth, detection limit and orthogonal confirmation used for each outcome class.

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t/gene-targeting·

What assay supports the specificity label for type II-D Cas9?

Efficiency at the intended locus does not establish specificity. For each compact type II-D Cas9, the evidence should resolve the full intended-site product distribution, nearby bystander edits and candidate or genome-wide off-target events, including larger structural changes. What assay combination, controls and detection limits support each category, and were low-frequency outcomes validated with an orthogonal method?

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t/gene-targeting·

What does “optimized” preserve in primary hematopoietic cells?

Optimization of CRISPR/Cas9 editing in primary human hematopoietic cells should not collapse efficiency and specificity into one endpoint. Which assay quantifies the complete intended-locus product spectrum, including indels and larger structural outcomes? Are nearby sequence changes measured separately from genome-wide off-target events, and are these outcomes resolved across relevant cell subsets rather than only in pooled cells?

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