Bacterial Artificial Chromosome Recombineering Core

The mission of the BAC Core is to provide access to a complex technology to investigators at the so that they can use their resources to conduct research instead of developing tools for research.

BACs contain large segments of chromosomal DNA (over 100,000 base pairs). The extensive DNA sequence contained in the BACs confers significant advantages to investigators who wish to use gene expression in their research. Shorter DNA fragments often do not contain enough gene expression information to recapitulate normal gene expression patterns (Giraldo and Montoliu, 2001). The BAC Core uses recombineering methods  to introduce genetic modifications in BACs by homologous recombination.

The BAC Core

  • purifies DNA from BAC clones for use in cell electroporation or the production of transgenic mice or rats
  • conducts pulsed field gel electrophoresis to restriction enzyme map BACs for purposes of identification
  • introduces reporter proteins such as green fluorescent proteins to mark cells
  • introduces point mutations in genes to modify the proteins produced in cells
  • introduces deletions into genes in the BAC to modifiy proteins or abrogate expression
  • knocks in cDNAs for expression for gene expression by heterlogous promoters
  • subclones BAC DNA fragments with gap repair plasmids
  • prepares gene targeting vectors for use in embryonic stem cells from BACs

BAC DNA purification methods

BAC DNA purification and Recombineering request forms can now be found on MiCores.

 

Examples of research projects that take advantage of BAC technology include establishment of transgenic mouse models of human diseases (Cui et al., 2000), and transgenic mouse experiments to identify homologs of human disease genes (Probst et al., 1998Wang et al., 1998). The Transgenic Core has a extensive experience in the production of BAC transgenic mice (Van Keuren et al., 2009) and transgenic rats so that the translation of genetically engineered BACs into genetically engineered mouse and rat models is straightforward.