Negative regulation of the RTBV promoter by designed zinc finger proteins.
The symptoms of rice tungro disease are caused by infection by a DNA-containing virus, rice tungro bacilliform virus (RTBV). To reduce expression of the RTBV promoter, and to ultimately reduce virus replication, we tested three synthetic zinc finger protein transcription factors (ZF-TFs), each comprised of six finger domains, designed to bind to sequences between -58 and +50 of the promoter. Two of these ZF-TFs reduced expression from the promoter in transient assays and in transgenic Arabidopsis thaliana plants. One of the ZF-TFs had significant effects on plant regeneration, apparently as a consequence of binding to multiple sites in the A. thaliana genome. Expression from the RTBV promoter was reduced by approximately 45% in transient assays and was reduced by up to 80% in transgenic plants. Co-expression of two different ZF-TFs did not further reduce expression of the promoter. These experiments suggest that ZF-TFs may be used to reduce replication of RTBV and thereby offer a potential method for control of an important crop disease.
Ordiz
M Isabel
MI
Donald Danforth Plant Science Center, 975 N. Warson Road, St. Louis, MO 63132, USA.
Magnenat
Laurent
L
Barbas
Carlos F
CF
Beachy
Roger N
RN
D017360:Arabidopsis / Q000235:genetics / Q000821:virology; D001483:Base Sequence; D015967:Gene Expression Regulation, Viral; D005966:Glucuronidase; D008969:Molecular Sequence Data; D012275:Oryza / Q000235:genetics / Q000821:virology; D010935:Plant Diseases / Q000821:virology*; D030821:Plants, Genetically Modified / Q000821:virology; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics*; D052641:Tungrovirus / Q000235:genetics*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics; D016335:Zinc Fingers
D012333:RNA, Messenger; D014157:Transcription Factors; D014764:Viral Proteins; D005966:Glucuronidase