Evidence map›Paper›PMID 41981915›Full record

ArticleMolecular therapy : the journal of the American Society of Gene Therapy2026

Messenger RNA and guide RNA distributions in lipid nanoparticles impact gene-editing efficiency in vivo.

Linh B Truong, Sixuan Li, Cosette Domkofski, Jinghan Lin, Carl Co, Sneha Halwasia, Michael Kuefner, Yizong Hu, Hai-Quan Mao, Tza-Huei Wang and 1 more

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Molecular therapy : the journal of the American Society of Gene Therapy, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing 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

2 citing papers in PubMed.

  1. Review
  2. 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

11 authors.

Linh B TruongArbor Biotechnologies, Cambridge, MA 02140, USA.
Sixuan LiDepartment of Mechanical Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA.
Cosette DomkofskiArbor Biotechnologies, Cambridge, MA 02140, USA.
Jinghan LinInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
Carl CoArbor Biotechnologies, Cambridge, MA 02140, USA.
Sneha HalwasiaArbor Biotechnologies, Cambridge, MA 02140, USA.
Michael KuefnerArbor Biotechnologies, Cambridge, MA 02140, USA.
Yizong HuInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21218, USA.
Hai-Quan MaoInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21218, USA; Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21218, USA; Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD 21218, USA. Electronic address: hmao@jhu.edu.
Tza-Huei WangDepartment of Mechanical Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21218, USA. Electronic address: thwang@jhu.edu.
Shengshuang ZhuArbor Biotechnologies, Cambridge, MA 02140, USA. Electronic address: lynzhu0105@gmail.com.

Funding

Development of a low-cost epigenetic screening assay for Pap specimen-based detection of early-stage ovarian cancer in high-risk womenR01CA260628 · NCI · JOHNS HOPKINS UNIVERSITY · PI PISANIC II, THOMAS RUSSELL, SHIH, IE-MING · 2021 to 2025
$2.1M
Integrated genotype-informed single-cell molecular antibacterial susceptibility testing platform for rapid and actionable diagnosis of bacteremiaR01AI183336 · NIAID · JOHNS HOPKINS UNIVERSITY · PI Tza-Huei Jeff Wang · 2024 to 2026
$2.1M
Engineered Lipid Nanoparticles and Microgel Matrix to Program Th1/Th2 Immune ResponseR01CA293906 · NCI · JOHNS HOPKINS UNIVERSITY · PI Hai-Quan Mao · 2025 to 2026
$1.2M
NCI NIH HHS R01 CA260628NCI NIH HHS R01 CA293906NIAID NIH HHS R01 AI183336
6 · The paper itself

Abstract

Lipid nanoparticles (LNPs) are among the most advanced non-viral vectors for CRISPR-based gene-editing therapeutics. Co-packaging of messenger RNA (mRNA) and guide RNA (gRNA) inherently produces heterogeneous payload distributions. The impact of this heterogeneity on editing performance remains unclear. Here, we utilize cylindrical illumination confocal spectroscopy (CICS) for single-particle interrogation of ALC-0315 and DLin-MC3-DMA LNPs prepared by three different mixing methods. CICS resolves four distinct subpopulations: co-encapsulated (50.7%-60.4%), gRNA only (30.0%-36.5%), mRNA only (2.0%-3.4%), and empty (4.2%-13.8%), and it uncovers broad, particle-to-particle variability in RNA copy number within each class. Structure-function analysis reveals that LNP formulation and mixing processes influence payload distribution, resulting in a negative correlation between the fraction of empty LNPs and RNA loading per particle. We further investigated the correlation between these quality attributes and therapeutic performance. In mice, ALC-0315 LNPs carrying higher cargo loads (9.8 vs. 8.0 mRNA copies and 25.4 vs. 20.3 gRNA copies per co-encapsulated particle) yielded 1.5-fold higher in vivo editing activity (55.4% vs. 36.3% insertions and deletions [indels]) despite nearly identical biophysical characteristics including LNP size and RNA encapsulation. These results establish payload distribution as a potential determinant of gene-editing potency and demonstrate single-particle CICS as a powerful tool for rational design of multi-component nucleic acid-delivery systems.

Indexed as

Gene EditingLipidsNanoparticlesRNA, Guide, CRISPR-Cas SystemsRNA, MessengerAnimalsCRISPR-Cas SystemsHumansLiposomesMiceLipid NanoparticlesLipidsLiposomesRNA, Guide, CRISPR-Cas SystemsRNA, Messengercomposition heterogeneitygene editinglipid nanoparticlesmRNA-gRNA co-deliverypayload distributionprocess-structure-function relationshiprational designsingle-particle analysis

Identifiers

PMID41981915
PMCPMC13242163

What OpenQuestion holds

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Read underepoch 390

Registered trials

None linked

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.