Furthermore, precise and efficient intermolecular integration of an incoming plasmid bearing attP into an established Epstein–Barr virus plasmid bearing attB was documented in human cells. This work is a demonstration of efficient, site-specific, unidirectional integration in mammalian cells.

Jun 10, 2024 · We transfected cells with PASSIGE components to integrate a 5.6-kb puromycin-encoding donor plasmid, containing a UMI, into the CCR5 locus via attB installation.

Co-transfecting cells with donor plasmid bearing attB site and helper plasmid encoding ΦC31 integrase results in integration at preintegrated wild-type attP or native cellular pseudo attP sites in the mammalian genomes.

Here we show that mutations in attB can block the integration reaction at different stages. Mutations at positions distal to the crossover site inhibit recombination by destabilizing the synapse with attP without significantly affecting DNA-binding affinity.

Oct 10, 2022 · LSRs canonically recombine two distinct DNA attachment sites natively found on an invading MGE (attP) and in the target bacterial genome (attB). Upon MGE insertion, the attachment sites are...

Oct 31, 2001 · Here we define compact attB and attP recognition sites for the R4 integrase and express the enzyme in mammalian cells. We demonstrate that R4 integrase functions in human cells, performing efficient and precise recombination between R4 attB and attP sites cloned on an extrachromosomal vector.

Apr 28, 2008 · φC31 integrase is a sequence-specific phage recombinase that can recombine two short DNA sequences called attB and attP. The enzyme can also promote genomic integration of plasmids carrying attB into native mammalian sequences having partial identity to attP.

Jan 11, 2025 · In molecular cloning, attP and attB sites are incorporated into vectors and host genomes to insert foreign DNA. Systems like Gateway cloning exploit these sites’ natural …

May 5, 2000 · Furthermore, precise and efficient intermolecular integration of an incoming plasmid bearing attP into an established Epstein–Barr virus plasmid bearing attB was documented in human cells. This work is a demonstration of efficient, site-specific, unidirectional integration in mammalian cells.

Jul 23, 2003 · Formation of the prophage state during lysogenic establishment typically involves the integration of the phage genome into a specific chromosomal attachment site (attB) in a particular orientation (Campbell et al., 1992; Campbell, 1992).