Cre Transgenic Mouse Lines
With the development of the Cre-LoxP system, conditional gene targeting has rapidly become a powerful technology that facilitates the study of gene function. This advanced technique circumvents three major concerns sometimes levelled against conventional transgenic and gene-targeting approaches. First of all, gene ablation may exert its effect in multiple cell and tissue types, creating a complex phenotype in which it is difficult to distinguish direct function in a particular tissue from secondary effects resulting from altered gene function in other tissues. Secondly, a gene deletion expressed in the germ line may cause embryonic lethality, thereby precluding analysis of gene function in the adult tissues. Thirdly, the transgenic approach represents a somewhat surreal over-expression of a given protein often causing spurious phenotypes. The generation of conditional knockout mice is a multiple-step process, which involves mating the flox mutant mouse line (essential exon/s of the gene of interest are flanked by two LoxP sites) and the Cre-expressing mouse line. Over the past few years many inducible and/or tissue-specific Cre mouse lines have been developed. This chapter will give a brief review of the generation of Cre-expressing mouse lines and will discuss the strategy of using these Cre lines. In addition, information regarding established Cre-expressing mouse lines will be provided.
- MacVector: Aligning Sequences
- Isolation of YAC Ends by Plasmid Rescue
- Long-Range PCR
- Two Flavors of Bulk Segregant Analysis in Yeast
- Combination of Native and Denaturing PAGE for the Detection of Protein Binding Regions in Long Fragments of Genomic DNA
- Fishn ChIPs: Chromatin Immunoprecipitation in the Zebrafish Embryo
- PAP-LMPCR: An Improved, Sequence-Selective Method for the In Vivo Analysis of Transcription Factor Occupancy and Chromatin Fine
- Inhibition of DNA Methylation in Somatic Cells
- Functional Expression of Type 1 Rat GABA Transporter in Microinjected Xenopus laevis Oocytes
- Direct Sequencing of PCR Products