Long-term BRCA1 down-regulation by small hairpin RNAs targeting the 3' untranslated region.
Mutations in the BRCA1 gene are responsible for the majority of hereditary breast/ovarian cancers. The functional significance of many mutations/splicing variants identified during the screening of high-risk individuals is difficult to predict due to the lack of in vitro functional tests correlating sequence variants with a risk of cancer development. RNA interference is a promising tool in analyzing functional properties of BRCA1 mutations. Here we designed and functionally analyzed shRNAs directed to 3'-UTR of BRCA1 mRNA that may be used to knock-down expression of endogenous BRCA1. Using retroviral infection, we achieved long-term down-regulation of BRCA1 in a cell-type specific manner. We propose that 3'-UTR-directed shRNAs, coupled with up-regulation of exogenous mutated BRCA1 variants, may constitute a versatile system for the functional analysis of BRCA1 gene alterations.
Vondruskova
E
E
Institute of Biochemistry and Experimental Oncology, Czech Republic. escho@lf1.cuni.cz
Malik
R
R
Sevcik
J
J
Kleiblova
P
P
Kleibl
Z
Z
D020413:3' Untranslated Regions / Q000235:genetics*; D045744:Cell Line, Tumor; D015536:Down-Regulation; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D019398:Genes, BRCA1 / Q000502:physiology; D006801:Humans; D034741:RNA, Small Interfering / Q000235:genetics*
D020413:3' Untranslated Regions; D034741:RNA, Small Interfering