Evidence map›Paper›PMID 38463141›Full record

ArticleMolecular therapy. Methods & clinical development2024

Thorough molecular configuration analysis of noncanonical AAV genomes in AAV vector preparations.

Junping Zhang, Xiangping Yu, Matthew Chrzanowski, Jiahe Tian, Derek Pouchnik, Ping Guo, Roland W Herzog, Weidong Xiao

Abstract read
In one paragraph

Article in Molecular therapy. Methods & clinical development, 2024. 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.

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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

8 authors.

Junping ZhangHerman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
Xiangping YuNikegen, Wynnewood, PA 19096, USA.
Matthew ChrzanowskiNikegen, Wynnewood, PA 19096, USA.
Jiahe TianDepartment of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
Derek PouchnikSchool of Molecular Biosciences, Washington State University, Pullman, WA 99164-4660, USA.
Ping GuoLewis Katz School of Medicine, Temple University, Philadelphia, PA 19140, USA.
Roland W HerzogHerman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
Weidong XiaoHerman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA.

Funding

Toward Safer Gene Therapy for Hemophilia AP01HL160472 · NHLBI · INDIANA UNIVERSITY INDIANAPOLIS · PI Roland W. Herzog · 2022 to 2026
$15.1M
Advancing CNS drug delivery via epigenetic modulationR01NS132504 · NINDS · CINCINNATI CHILDRENS HOSP MED CTR · PI Dao Pan · 2023 to 2026
$2.5M
Novel adeno-associated virus vector production system developmentR01HL114152 · NHLBI · TEMPLE UNIV OF THE COMMONWEALTH · PI XIAO, WEIDONG · 2012 to 2016
$2.2M
PURPOSE OF THE NGVB CONTRACT IS TO CONTINUE SUPPORTING GENE THERAPY RESEARCH.75N92019D00018 · OD · INDIANA UNIVERSITY INDIANAPOLIS · 2019 to 2023
$1.7M
REGULATION OF THE BIOSYNTHESIS OF THE OPIOID PEPTIDES AND OTHER NEUROPEPTIDESZ01HL000018 · NHLBI · HEART, LUNG, AND BLOOD INSTITUTE · PI SABOL, S L · 1985 to 1992
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NHLBI NIH HHS 75N92019D00018NHLBI NIH HHS P01 HL160472NHLBI NIH HHS R01 HL114152NINDS NIH HHS R01 NS132504
6 · The paper itself

Abstract

The unique palindromic inverted terminal repeats (ITRs) and single-stranded nature of adeno-associated virus (AAV) DNA are major hurdles to current sequencing technologies. Due to these characteristics, sequencing noncanonical AAV genomes present in AAV vector preparations remains challenging. To address this limitation, we developed thorough molecule configuration analysis of noncanonical AAV genomes (TMCA-AAV-seq). TMCA-AAV-seq takes advantage of the documented AAV packaging mechanism in which encapsidation initiates from its 3' ITR, for AAV-seq library construction. Any AAV genome with a 3' ITR is converted to a template suitable to adapter addition by a Bst DNA polymerase-mediated extension reaction. This extension reaction helps fix ITR heterogeneity in the AAV population and allows efficient adapter addition to even noncanonical AAV genomes. The resulting library maintains the original AAV genome configurations without introducing undesired changes. Subsequently, long-read sequencing can be performed by the Pacific Biosciences (PacBio) single-molecule, real-time (SMRT) sequencing technology platform. Finally, through comprehensive data analysis, we can recover canonical, noncanonical AAV DNA, and non-AAV vector DNA sequences, along with their molecular configurations. Our method is a robust tool for profiling thorough AAV-population genomes. TMCA-AAVseq can be further extended to all parvoviruses and their derivative vectors.

Indexed as

AAVadeno-associated viruslibrary constructionmolecular configurationnoncanonical AAV genomesTMCA-AAVseq

Identifiers

PMID38463141
PMCPMC10924063

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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.