Abstract
RNA interference (RNAi) is a naturally occurring posttranscriptional gene-silencing mechanism that has been adapted as a genetic tool for loss-of-function studies of a variety of organisms. It is more widely applicable than classical gene targeting and allows for the simultaneous inactivation of several homologous genes with a single transgene. Recently, RNAi has been used for conditional and conventional gene inactivation in mice. Unlike gene targeting, RNAi is a dynamic process, and its efficiency may vary both between cell types and throughout development. Here we demonstrate that RNAi can be used to target three separately encoded isoforms of the bcl-2 family gene bfl-1/A1 in a conditional manner in mice. The extent of gene inactivation varies between different cell types and is least efficient in mature lymphocytes. Our data suggest that RNAi is affected by factors beyond small interfering RNA-mRNA stoichiometry.
Publication types
-
Research Support, N.I.H., Extramural
-
Research Support, Non-U.S. Gov't
-
Research Support, U.S. Gov't, P.H.S.
MeSH terms
-
Animals
-
Cell Differentiation
-
Cells, Cultured
-
Gene Expression Regulation, Developmental*
-
Hematopoietic System / metabolism*
-
Integrases / genetics
-
Integrases / metabolism
-
Mice
-
Organ Specificity
-
Peptide Initiation Factors / genetics
-
Promoter Regions, Genetic / genetics*
-
Proto-Oncogene Proteins c-bcl-2 / genetics
-
RNA Interference*
-
RNA, Messenger / genetics
-
RNA, Messenger / metabolism
-
RNA, Small Interfering / genetics
-
RNA, Small Interfering / metabolism*
-
RNA, Small Nuclear / genetics*
-
T-Lymphocytes / cytology
-
T-Lymphocytes / metabolism
-
Thymus Gland / cytology
-
Thymus Gland / metabolism
-
Transgenes / genetics*
-
Viral Proteins / genetics
-
Viral Proteins / metabolism
Substances
-
Peptide Initiation Factors
-
Proto-Oncogene Proteins c-bcl-2
-
RNA, Messenger
-
RNA, Small Interfering
-
RNA, Small Nuclear
-
U6 small nuclear RNA
-
Viral Proteins
-
Cre recombinase
-
Integrases