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Genesis March 1, 2012; 50 (3): 307-15.

Simple, fast, tissue-specific bacterial artificial chromosome transgenesis in Xenopus.

Fish MB , Nakayama T , Grainger RM .

We have developed a method of injecting bacterial artificial chromosome (BAC) DNA into Xenopus embryos that is simple and efficient, and results in consistent and tissue-specific expression of transgenes cloned into BAC vectors. Working with large pieces of DNA, as can be accommodated by BACs, is necessary when studying large or complex genes and conducive to studying the function of long-range regulatory elements that act to control developmentally restricted gene expression. We recombineered fluorescent reporters into three Xenopus tropicalis BAC clones targeting three different genes and report that up to 60% of injected embryos express the reporter in a manner consistent with endogenous expression. The behavior of these BACs, which are replicated after injection, contrasts with that of smaller plasmids, which degrade relatively quickly when injected as circular molecules and generally fail to recapitulate endogenous expression when not integrated into the Xenopus genome.

PubMed ID: 22084035
PMC ID: PMC3870158
Article link: Genesis
Grant support: EY017400 NEI NIH HHS , EY019000 NEI NIH HHS , R01 EY017400 NEI NIH HHS , R01 EY018000 NEI NIH HHS , R43 EY019000 NEI NIH HHS , R44 EY019000 NEI NIH HHS

Genes referenced: atoh7 pax6 rax

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Andreazzoli, 2003, Pubmed, Xenbase [+]

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