ArticleNature biotechnology2026
A pervasive RT-qPCR artifact inflates RNA knockdown by RNA-targeting CRISPR.
Article in Nature biotechnology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
3 citing papers in PubMed.
- Architecture of a DNA-guided Cas12a.Nature structural & molecular biology · 2026Article
- From reading to writing.Nature biotechnology · 2026Article
- Guide DNA - not RNA - expands the CRISPR toolkit.Nature biotechnology · 2026Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
3 authors.
Funding
Abstract
Knockdown efficiency of RNA-targeting CRISPR systems is commonly measured by quantitative polymerase chain reaction with reverse transcription (RT-qPCR). Here we discovered that guide RNAs copurify during RNA extraction and inhibit RT-qPCR for amplicons spanning or upstream of the guide RNA binding site, resulting in overestimation of knockdown efficiency across all CRISPR systems tested. We recommend using a processive reverse transcriptase with strong strand-displacing activity, with orthogonal methods, to ensure accurate quantification when using RT-qPCR.
Identifiers
42680825What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.