Mutagenesis by reversible promoter insertion to study the activation of NF-kappaB.

ABSTRACT: Genetic dissection of signaling pathways in mammalian cells involves screening or selecting phenotypic mutants obtained by a variety of techniques. Limitations in current methods include inadequate genome coverage and difficulty in validating the link between mutation and phenotype. We describe an improved method for insertional mutagenesis with retroviral vectors and show that the ability to induce mutations increases greatly if a randomly inserted promoter directs transcription into the host DNA. The mutant phenotype is due to the expression of a hybrid transcript derived from the vector and the insertion site. Because other alleles of the affected gene remain intact, the phenotype is dominant, but is reversible by inactivating the promoter, for example, by site-specific recombination. I

SUBMITTER: Kandel ES 

PROVIDER: S-EPMC1088393 | biostudies-literature | 2005 May

REPOSITORIES: biostudies-literature