Limitations
What this work cannot tell you.
This is not a disclaimer page. It is a list of the specific boundary of each claim on this site, which is the part most easily lost when work gets summarised.
Nothing here was measured in a heart
Every functional number on this site comes from cells grown in a dish and engineered to produce this channel. That is the standard way to measure a variant, and it is not a heart. A heart has different cell types, different partner proteins, a different environment, and a gradient across the ventricular wall that no dish reproduces.
Applies to: the 68.3 percent figure, the 31.3 percent rescaling, and every statement about current.
The central mechanistic question is open
Whether the broken copy simply fails or actively interferes with the working copy has not been resolved. The standard assay cannot resolve it: the two models predict the same number. Any statement on this site about what a therapy would do rests on that unresolved question, and would need revisiting if the answer came out the other way.
Computational prediction nominates, it does not prove
The base-editing guide, the off-target scan, and the variant nominations are predictions. A predicted off-target site is a place worth checking, not a place where editing was observed. A nominated variant is a candidate for testing, not a variant known to be harmful.
This project has direct evidence of how badly prediction can fail here: given four variants already known to break the channel, the folding-stability method it had been relying on identified one.
Where this project's own numbers disagree
Several quantities have been measured more than once here, by different methods, and do not agree. Where that is true no single value is quoted on this site.
- How buried position 104 is inside the assembled channel has five different values across five analyses, depending on the method and on whether the domain is measured alone or in the whole protein.
- The count of protein-changing off-target sites for the lead editing guide was revised upward when a more thorough search was run. The revised figure is the one that stands, and the earlier one still appears in places.
- The nearest protein-changing off-target moved closer once alignments with an inserted or deleted letter were modelled. The earlier statement that the nearest sat at three mismatches is no longer true.
These are listed rather than reconciled silently, because a project that quietly harmonises its own disagreements is one you cannot check.
Specific boundaries, by result
| The result | What it does not establish |
|---|---|
| Off-target scan for the base-editing guide | It was run to a set number of mismatches, and the tail beyond that was not annotated. Bulged alignments were not modelled. Complete within its stated span, not absolutely complete. |
| The upregulation ceiling | It rules out one specific mechanism, boosting output by redirecting non-productive message. It does not rule out every way of raising channel output. |
| Seventeen nominated variants | They are candidates for testing. The other 114 are uninformative rather than benign, which is a different thing and matters to anyone carrying one. |
| The reclassification argument | It is an argument that the evidence should be reviewed. Only the submitting laboratories and the expert panel can change a classification. |
| The structural argument | One deposited structure, one conformation. Proteins move, and a single set of coordinates does not capture that. |
Work that was retired
An earlier version of this site presented a different hypothesis: that a small molecule could stabilise the protein while it folded, and it carried a screen of candidate compounds. That work has been retired. The prediction method underneath it failed its own calibration, and the route carries a hazard if the broken copy turns out to interfere with the working one.
It is named here rather than quietly deleted, because anyone who read the earlier version deserves to know it was withdrawn and why. Nothing from that campaign should be cited.
Not peer reviewed
None of these results has been through peer review. They are posted as preprints so the reasoning is visible early. That is a deliberate choice, and it means the appropriate level of confidence is provisional.