Evidence map›Paper›PMID 37521788›Full record

ArticleACS central science2023

Spacer Fidelity Assessments of Guide RNA by Top-Down Mass Spectrometry.

Luis A Macias, Sara P Garcia, Kayla M Back, Yue Wu, G Hall Johnson, Sekar Kathiresan, Andrew M Bellinger, Ellen Rohde, Michael A Freitas, James A Madsen

Abstract read
In one paragraph

Article in ACS central science, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.

0numbers the graph read from it
0cells of the map it votes in
10citing 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

10 citing papers in PubMed.

  1. Article
  2. Article
  3. Article
  4. Article
  5. Article
  6. Article
  7. Article
  8. Development and IND-enabling studies of a novel Cas9 genome-edited autologous CD34Molecular therapy : the journal of the American Society of Gene Therapy · 2024
    Article
  9. Article
  10. Review
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

10 authors.

Luis A MaciasVerve Therapeutics, 201 Brookline Avenue, Suite 601, Boston, Massachusetts 02215, United States.ORCID https://orcid.org/0000-0003-1949-6029
Sara P GarciaVerve Therapeutics, 201 Brookline Avenue, Suite 601, Boston, Massachusetts 02215, United States.
Kayla M BackVerve Therapeutics, 201 Brookline Avenue, Suite 601, Boston, Massachusetts 02215, United States.
Yue WuVerve Therapeutics, 201 Brookline Avenue, Suite 601, Boston, Massachusetts 02215, United States.
G Hall JohnsonMassMatrix, Inc., 600 Teteridge Road, Columbus, Ohio 43214, United States.
Sekar KathiresanVerve Therapeutics, 201 Brookline Avenue, Suite 601, Boston, Massachusetts 02215, United States.
Andrew M BellingerVerve Therapeutics, 201 Brookline Avenue, Suite 601, Boston, Massachusetts 02215, United States.
Ellen RohdeVerve Therapeutics, 201 Brookline Avenue, Suite 601, Boston, Massachusetts 02215, United States.
Michael A FreitasMassMatrix, Inc., 600 Teteridge Road, Columbus, Ohio 43214, United States.ORCID https://orcid.org/0000-0002-3103-8410
James A MadsenVerve Therapeutics, 201 Brookline Avenue, Suite 601, Boston, Massachusetts 02215, United States.ORCID https://orcid.org/0009-0009-2512-8944

Funding

Accelerating Gene Therapy and Editing with Advanced MS-Based Data Analysis for Nonstandard and Hybrid Nucleotide SequencesR44GM152984 · NIGMS · MASSMATRIX, INC. · PI JOHNSON, GEORGE HALL · 2023 to 2024
$1.7M
Accelerating Site-specific Characterization of Protein Therapeutics with Novel Machine Learning MethodsR44GM131484 · NIGMS · MASSMATRIX, INC. · PI JOHNSON, GEORGE HALL · 2019 to 2021
$1.1M
NIGMS NIH HHS R44 GM131484NIGMS NIH HHS R44 GM152984
6 · The paper itself

Abstract

The advancement of CRISPR-based gene editing tools into biotherapeutics offers the potential for cures to genetic disorders and for new treatment paradigms for even common diseases. Arguably, the most important component of a CRISPR-based medicine is the guide RNA, which is generally large (>100-mer) synthetic RNA composed of a "tracr" and "spacer" region, the latter of which dictates the on-target editing site as well as potential undesired off-target edits. Aiming to advance contemporary capabilities for gRNA characterization to ensure the spacer region is of high fidelity, top-down mass spectrometry was herein implemented to provide direct and quantitative assessments of highly modified gRNA. In addition to sequencing the spacer region and pinpointing modifications, top-down mass spectra were utilized to quantify single-base spacer substitution impurities down to <1% and to decipher highly dissimilar spacers. To accomplish these results in an automated fashion, we devised custom software capable of sequencing and quantifying impurities in gRNA spacers. Notably, we developed automated tools that enabled the quantification of single-base substitutions, including advanced isotopic pattern matching for C > U and U > C substitutions, and created a

Identifiers

PMID37521788
PMCPMC10375574

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LicenceCC BY
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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.