title: T7-RNA polymerase dependent RNAi system in Aspergillus fumigatus: a proof of concept study.
abstract: An RNAi system based on T7 RNA polymerase (TRNAP) was designed and examined in Aspergillus fumigatus. This system consists of two elements; an inducible T7RNAP expressing cassette and an AMA1-based episomal RNAi plasmid. These constructs were transformed into the A. fumigatus protoplasts and the efficiency of this system was tested in downregulation of alb1 gene. Upon the induction of T7RNAP expression, the recombinant T7RNAP was able to recognize T7 promoters, which were located on the episomal plasmid and in opposite direction. As a result, the bidirectional transcription of alb1 fragment led to the silencing of the target gene. However, our results demonstrated that this silencing system is unstable and may not be applicable in preparation of RNAi libraries.
authors: Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Ave, 1316943551 Tehran, Iran.
authors: Somayeh
authors: S
authors: Enayati
authors: Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Ave, 1316943551 Tehran, Iran.
authors: Mohammad
authors: M
authors: Azizi
authors: Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Ave, 1316943551 Tehran, Iran.
authors: Elahe
authors: E
authors: Aminollahi
authors: Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Ave, 1316943551 Tehran, Iran.
authors: Mona
authors: M
authors: Ranjvar Shahrivar
authors: Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Ave, 1316943551 Tehran, Iran v_khalaj@yahoo.com.
authors: Vahid
authors: V
authors: Khalaj
mesh_terms: D001232:Aspergillus fumigatus / Q000235:genetics*; D064199:Cdc20 Proteins / Q000235:genetics*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D014170:Transformation, Genetic / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D064199:Cdc20 Proteins; D034741:RNA, Small Interfering; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases
keywords: Aspergillus fumigatus; RNAi; T7RNA polymerase