Evidence map›Paper›PMID 28654067›Full record

ArticleJournal of visualized experiments : JoVE2017

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow.

Gajendra W Suryawanshi, Song Xu, Yiming Xie, Tom Chou, Namshin Kim, Irvin S Y Chen, Sanggu Kim

Abstract readVideo-Audio Media
In one paragraph

Article in Journal of visualized experiments : JoVE, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing 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

4 citing papers in PubMed.

  1. Article
  2. Article
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  4. 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

7 authors.

Gajendra W SuryawanshiUCLA AIDS Institute, University of California at Los Angeles (UCLA); Department of Microbiology, Immunology, & Molecular Genetics, University of California at Los Angeles (UCLA).
Song XuDepartments of Biomathematics and Mathematics, University of California at Los Angeles (UCLA).
Yiming XieUCLA AIDS Institute, University of California at Los Angeles (UCLA).
Tom ChouDepartments of Biomathematics and Mathematics, University of California at Los Angeles (UCLA).
Namshin KimPersonalized Genomic Medicine Research Center, Division of Strategic Research Groups, Korea Research Institute of Bioscience and Biotechnology.
Irvin S Y ChenUCLA AIDS Institute, University of California at Los Angeles (UCLA); Department of Medicine, University of California at Los Angeles (UCLA); syuchen@mednet.ucla.edu.
Sanggu KimDepartment of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University (OSU); kim.6477@osu.edu.

Funding

Virology CoreP30AI028697 · NIAID · UNIVERSITY OF CALIFORNIA LOS ANGELES · PI KITCHEN, SCOTT G · 1991 to 2018
$39.9M
Viral Immunology CoreU19AI117941 · NIAID · UNIVERSITY OF CALIFORNIA LOS ANGELES · PI SYMONDS, GEOFFREY · 2015 to 2019
$10.6M
Defining a therapeutic threshold in HSPC gene therapy for HIV-1 diseaseR00HL116234 · NHLBI · OHIO STATE UNIVERSITY · PI KIM, SANGGU · 2016 to 2018
$747k
Mouse and mathematical models for HIV-1 suppression through HSPCR56HL126544 · NHLBI · UNIVERSITY OF CALIFORNIA LOS ANGELES · PI CHEN, IRVIN S.Y., CHOU, TOM · 2014 to 2014
$381k
NHLBI NIH HHS R00 HL116234NHLBI NIH HHS R56 HL126544NIAID NIH HHS P30 AI028697NIAID NIH HHS U19 AI117941
6 · The paper itself

Abstract

Integration Site (IS) assays are a critical component of the study of retroviral integration sites and their biological significance. In recent retroviral gene therapy studies, IS assays, in combination with next-generation sequencing, have been used as a cell-tracking tool to characterize clonal stem cell populations sharing the same IS. For the accurate comparison of repopulating stem cell clones within and across different samples, the detection sensitivity, data reproducibility, and high-throughput capacity of the assay are among the most important assay qualities. This work provides a detailed protocol and data analysis workflow for bidirectional IS analysis. The bidirectional assay can simultaneously sequence both upstream and downstream vector-host junctions. Compared to conventional unidirectional IS sequencing approaches, the bidirectional approach significantly improves IS detection rates and the characterization of integration events at both ends of the target DNA. The data analysis pipeline described here accurately identifies and enumerates identical IS sequences through multiple steps of comparison that map IS sequences onto the reference genome and determine sequencing errors. Using an optimized assay procedure, we have recently published the detailed repopulation patterns of thousands of Hematopoietic Stem Cell (HSC) clones following transplant in rhesus macaques, demonstrating for the first time the precise time point of HSC repopulation and the functional heterogeneity of HSCs in the primate system. The following protocol describes the step-by-step experimental procedure and data analysis workflow that accurately identifies and quantifies identical IS sequences.

Indexed as

WorkflowAnimalsCells, CulturedClone CellsGenetic VectorsHematopoietic Stem CellsHigh-Throughput Nucleotide SequencingPolymerase Chain ReactionReproducibility of ResultsRetroviridaeSequence Analysis, DNAVirus Integration

Identifiers

PMID28654067
PMCPMC5608418

What OpenQuestion holds

Textmetadata
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.