David Weinberg14 Billings St, Boston, MA 02132

2012030, Dec 6, 2012

Sep 10, 2010

13/394834

David P. Bartel - Brookline MA, US
Ines A. Drinnenberg - Cambridge MA, US
David E. Weinberg - Cambridge MA, US
Kathleen T. Xie - North Billerica MA, US
Kenneth H. Wolfe - Dublin, IE
Gerald Fink - Cambridge MA, US

Whitehead Institute for Biomedical Research - Cambridge MA
of Queen Elizabeth Near Dublin - Dublin

C12N 1/19
C12N 9/22
C12N 15/52

435199, 4352542, 536 232

The invention provides budding yeast that have a functional RNAi pathway. The invention provides RNAi pathway polypeptides derived from budding yeast that have an endogenous RNAi pathway. In some embodiments the invention provides functional budding yeast Dicer polypeptides and variants thereof. In some embodiments the invention provides functional budding yeast Argonaute polypeptides and variants thereof. Also provided are isolated nucleic acids encoding the polypeptides of the invention, vectors comprising such nucleic acids, and methods of making the polypeptides and nucleic acids. The invention further provides genetically engineered cells that comprise a functional RNAi pathway polypeptide derived from budding yeast. In some embodiments such cells lack a functional endogenous RNAi pathway and are genetically engineered to have a functional RNAi pathway by introducing nucleic acid(s) encoding one or more functional RNAi pathway polypeptides derived from budding yeast. The invention provides methods of using RNAi in budding yeast and/or in cells of other types, wherein the cells have been genetically engineered to express one or more RNAi pathway polypeptides of the invention. Also provided are methods of producing siRNA, either in vitro or in vivo, using a Dicer polypeptide derived from budding yeast.