Related Experiment Video
Updated: Feb 10, 2026
09:31
Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
Published on: March 22, 2016
18.4K
Screen and Verification for Transgene Integration Sites in Pigs
Linyuan Ma1, Yuzhe Wang1, Haitao Wang1
1The State Key Laboratory for Agricultural Biotechnology, College of Biological Science, China Agricultural University, Beijing, 100193, China.
Scientific Reports
|May 11, 2018
Summary
Researchers identified Pifs501 as a safe and effective site for transgene insertion in pigs, enabling efficient gene expression without disrupting nearby genes. This breakthrough advances pig genome engineering for gene therapy applications.
Area of Science:
- Genomics
- Biotechnology
- Gene Therapy
Background:
- Efficient transgene expression is crucial for gene therapy, but random integration poses risks.
- Identifying specific, safe genomic loci for transgene insertion is essential for predictable outcomes.
Purpose of the Study:
- To develop and validate a method for identifying optimal transgene integration sites in the pig genome.
- To establish a versatile cell line for precise gene insertion and modification in pigs.
Main Methods:
- In silico screening of the pig genome to identify candidate integration sites (Pifs302, Pifs501).
- CRISPR/Cas9-mediated site-specific integration of EGFP at Pifs501.
- Functional verification through in vitro and in vivo assays in transgenic pigs.
- Epigenetic analysis (hypomethylation) of the Pifs501 locus.
Main Results:
- Pifs501 was identified as an actively transcribed intergenic region with favorable epigenetic properties.
- High and ubiquitous EGFP expression was achieved in various pig cell types and tissues post-integration.
- No adverse effects on neighboring gene expression (600kb) were observed.
- A master cell line at Pifs501 enabled seamless EGFP to Follistatin exchange using Cre/loxP.
Conclusions:
- The Pifs501 locus is a highly effective and safe site for transgene insertion in pigs.
- The combined bioinformatics and functional verification strategy is validated for identifying genomic safe harbors.
- Pifs501 offers significant potential for advancing pig genome engineering and gene therapy applications.
Related Concept Videos
03:00Self-Evaluation: Self-Enhancement and Self-Verification
5.8K
Social psychologists have documented that feeling good about ourselves and maintaining positive self-esteem is a powerful motivator of human behavior (Tavris & Aronson, 2008). In the United States, members of the predominant culture typically think very highly of themselves and view themselves as good people who are above average on many desirable traits (Ehrlinger, Gilovich, & Ross, 2005). Often, our behavior, attitudes, and beliefs are affected when we experience a threat to our...
5.8K
01:17Strategies of Self-Presentation II: Self-Verification
204
Self-verification is a fundamental psychological drive wherein individuals seek affirmation of their self-concept from others, striving for consistency between their internal self-view and external perceptions. This drive operates even when the self-concept is negative, influencing interpersonal behavior and feedback preferences in complex and often counterintuitive ways. Unlike the self-enhancement motive, which seeks positive evaluations, self-verification prioritizes coherence and...
204
00:53Transgenic Organisms
33.6K
Overview
33.6K
01:49Conserved Binding Sites
5.2K
Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...
5.2K
01:49Conserved Binding Sites
2.0K
2.0K
02:40Ligand Binding Sites
15.2K
Proteins are dynamic macromolecules that carry out a wide variety of essential processes; however, the activities of most proteins depend on their interactions with other molecules or ions, known as ligands.
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...
15.2K