ArticleMethods in molecular biology (Clifton, N.J.)2025
Identifying Genomic DNA Sequences Near the Nuclear Lamina Using Proximity Biotinylation with Ascorbate Peroxidase.
Article in Methods in molecular biology (Clifton, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Ascorbate peroxidase (APEX2) is an engineered plant-based enzyme that is used for the proximal ligation of biotin onto nearby proteins and nucleic acids. The APEX2 enzyme can be targeted to a specific subcellular compartment (e.g., the nuclear lamina) by creating a fusion between the APEX2 enzyme and a targeting protein (e.g., Lamin B1). We have found that the APEX2 proximity ligation reaction is very robust and can be performed in both living and formaldehyde-fixed cells. The biotinylated proteins and nucleic acids can then be precipitated with streptavidin beads for the identification of proximal proteins, RNA, and DNA. Here, we detail a protocol for the use of an APEX-Lamin B1 fusion protein to identify DNA in the proximity of the nuclear lamina, including lamina-associated domains (LADs).
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