Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Mapping the zygote-to-adult developmental cell phylogeny in Arabidopsis thaliana reveals a three-cell rule of branching.

Nature plants·2026
Same author

Author Correction: Split complementation of base editors to minimize off-target edits.

Nature plants·2024
Same author

Split complementation of base editors to minimize off-target edits.

Nature plants·2023
Same author

After Skin Wounding, Noncoding dsRNA Coordinates Prostaglandins and Wnts to Promote Regeneration.

The Journal of investigative dermatology·2017
Same author

Metabolic profiling and novel plasma biomarkers for predicting survival in epithelial ovarian cancer.

Oncotarget·2017
Same author

Identification of a six-lncRNA signature associated with recurrence of ovarian cancer.

Scientific reports·2017

Related Experiment Video

Updated: Sep 12, 2025

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow
12:53

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow

Published on: June 14, 2017

10.9K

Genome-wide profiling the integration patterns with T7-PCR.

Ang Li1, Chichen Xiao1, Jinwen Wang1

  • 1State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, China.

The Plant Journal : for Cell and Molecular Biology
|August 9, 2025
PubMed
Summary
This summary is machine-generated.

We developed T7-PCR, a novel method for accurately mapping gene integration sites. This technique enhances signal-to-noise ratio, improving efficiency and accuracy for genome engineering applications.

Keywords:
genome engineeringmapping insertion sitestargeted integration

More Related Videos

Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
09:31

Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites

Published on: March 22, 2016

17.8K
Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector
12:08

Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector

Published on: March 28, 2018

12.6K

Related Experiment Videos

Last Updated: Sep 12, 2025

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow
12:53

Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow

Published on: June 14, 2017

10.9K
Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
09:31

Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites

Published on: March 22, 2016

17.8K
Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector
12:08

Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector

Published on: March 28, 2018

12.6K

Area of Science:

  • Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • Gene integration is crucial for gene function studies, molecular breeding, and gene therapy.
  • Accurate identification of integration sites is vital for safety and efficacy in genome engineering.
  • Current mapping methods face challenges with high costs and low signal-to-noise ratios.

Purpose of the Study:

  • To develop an innovative and efficient tool for mapping exogenous gene integration sites.
  • To overcome the limitations of existing high-cost, low-signal-to-noise ratio mapping techniques.
  • To enhance the accuracy and efficiency of integration site identification in various cell types.

Main Methods:

  • Developed T7 polymerase-mediated in vitro transcription (T7-IVT) for capturing junction fragments.
  • Converted genomic flanking sequences into RNA to enrich junction fragments and reduce background DNA.
  • Named and validated the T7-PCR method across yeast, plant, and human cells.

Main Results:

  • T7-PCR significantly enhances the signal-to-noise ratio by enriching junction fragments and eliminating background DNA.
  • The method demonstrated high efficiency and accuracy in diverse cell types and integration scenarios.
  • T7-PCR showed minimal positional effects, outperforming current next-generation sequencing (NGS)-based strategies.

Conclusions:

  • T7-PCR offers a cost-effective and accurate solution for mapping gene integration sites.
  • This method is highly applicable for high-throughput transgene screening.
  • T7-PCR supports the development of next-generation tools for large fragment targeted integration.