GENETIC TARGETING IN NON-CONVENTIONAL YEAST USING AN RNA-GUIDED ENDONUCLEASE

摘要

Non-conventional yeasts are disclosed herein comprising at least one RNA-guided endonuclease (RGEN) comprising at least one RNA component that does not have a 5'-cap. This uncapped RNA component comprises a sequence complementary to a target site sequence in a chromosome or episome in the yeast. The RGEN can bind to, and optionally cleave, one or both DNA strands at the target site sequence. An example of an RGEN herein is a complex of a Cas9 protein with a guide RNA. A ribozyme is used in certain embodiments to provide an RNA component lacking a 5'-cap. Further disclosed are methods of genetic targeting in non-conventional yeast.