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Updated: May 3, 2026
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Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
Published on: March 22, 2016
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An in vitro transcription-based protocol for mapping HIV integration sites using lentiviral integration site
Kamil Więcek1, Janusz Wiśniewski1, Heng-Chang Chen1
1Quantitative Virology Research Group, Population Diagnostics Center, Łukasiewicz Research Network - PORT Polish Center for Technology Development, Stablowicka 147, 54-066 Wrocław, Poland.
STAR Protocols
|December 23, 2025
Summary
We developed a rapid method called lentiviral integration site sequencing (LIS-seq) to map human immunodeficiency virus (HIV) integration sites. This technique aids in creating new antiretroviral therapies.
Area of Science:
- Virology
- Genomics
- Molecular Biology
Background:
- Identifying human immunodeficiency virus (HIV) integration sites is critical for designing effective antiretroviral strategies.
- Current methods for mapping integration sites can be time-consuming and complex.
Purpose of the Study:
- To present a novel, rapid, in vitro transcription-based method for lentiviral integration site sequencing (LIS-seq).
- To enable efficient and accurate mapping of HIV integration sites in cellular models.
Main Methods:
- Establishment of clonal cellular models infected with an HIV-based vector.
- Genomic DNA isolation, fragmentation, and T7 in vitro transcription.
- Poly(A) tailing, sequencing library generation, and bioinformatic analysis pipeline.
Main Results:
- Demonstrated the successful application of LIS-seq to clonal cellular models.
- LIS-seq enables rapid and precise identification of lentiviral integration sites.
Conclusions:
- LIS-seq provides a streamlined approach for integration site analysis.
- This method facilitates the development of advanced antiretroviral therapies by understanding viral integration patterns.
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