Evidence map›Paper›PMID 41674724›Full record

ArticleMolecular therapy. Nucleic acids2026

CRISPR-MiX: A pooled single-stranded donor strategy to enhance HDR efficiency in human iPSCs.

Rachel Baum, Narasimha Telugu, Arne A N Bruyneel, Maryam Kay, Pooja Nair, Isaac Perea-Gil, Vittavat Termglinchan, Nike Bharucha, Edina Lee, Mark Mercola and 2 more

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Molecular therapy. Nucleic acids, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

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.

2 · The registry

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.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

12 authors.

Rachel BaumCardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
Narasimha TeluguMax Delbrück Center for Molecular Medicine (MDC), 13125 Berlin, Germany.
Arne A N BruyneelCardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
Maryam KayCardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
Pooja NairCardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
Isaac Perea-GilCardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
Vittavat TermglinchanCardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
Nike BharuchaCardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
Edina LeeCardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
Mark MercolaCardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
Sebastian DieckeMax Delbrück Center for Molecular Medicine (MDC), 13125 Berlin, Germany.
Ioannis KarakikesCardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

CRISPR-Cas9 is widely used to model genetic disorders by introducing or correcting disease-associated mutations in induced pluripotent stem cells (iPSCs) through homology-directed repair (HDR). However, HDR efficiency in iPSCs remains low and is highly dependent on the target locus. Here, we developed CRISPR-MiX, an improved protocol to enhance HDR efficiency in human iPSCs. Using a GFP-to-BFP reporter system, we identified key single-stranded oligodeoxynucleotide (ssODN) donor design parameters, including homology arm symmetry, CRISPR/Cas-blocking mutations, and strand complementarity, which significantly influence HDR outcomes. We applied this approach to introduce pathogenic variants into five genes related to genetic cardiomyopathies. Quantitative analysis of HDR events showed that both the target locus and ssODN design strongly affect HDR efficiency. To address the locus- and design-specific limitations, we established CRISPR-MiX, a pooled ssODN-based method for scarless genome editing using ribonucleoproteins (RNPs) that does not require selection. CRISPR-MiX consistently improved HDR efficiency across multiple loci. This strategy offers a simple, robust, and versatile approach for precise genome engineering in iPSCs, supporting broad applications in disease modeling and functional genomics.

Indexed as

cardiomyopathyCRISPR-Cas9disease modelingDNA breaksDNA repairgenome editinghomology-directed repairiPSCsMT: RNA/DNA Editingpluripotent stem cellsssODN

Identifiers

PMID41674724
PMCPMC12887267

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Registered trials

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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.