Engineered microbial population

Abstract

Provided herein are genetically engineered bacteria that are native to a host insect microbiome. Further provided are methods of inducing RNA interference in an insect, such as a bee, by administering the genetically engineered bacteria.

1 - 17 . (canceled) 
     
     
         18 . A method for downregulating expression of a target gene product, comprising administering an effective amount of a composition comprising one or more bacteria genetically engineered to express at least one heterologous nucleic acid, wherein the one or more bacteria are native to the microbiome of a host insect to an insect, wherein said bacteria express an inhibitor of said target gene product. 
     
     
         19 . The method of  claim 18 , wherein the target gene product is a gene from an organism selected from the group consisting of a pathogen, a parasite, a virus, a mite, Acute Bee Paralysis Virus (ABPV), Kashmir Bee Virus (KBV), Israeli Acute Paralysis Virus (IAPV),  Nosema ceranae , Deformed Wing Virus,  Varroa destructor  mite, and Small Hive Beetle. 
     
     
         20 . The method of  claim 18 , wherein the insect is selected from the group consisting of a bee, a honey bee, a forager, a hive bee, a pupae, an adult bee, and a bee colony parasite. 
     
     
         21 . The method of  claim 18 , wherein the target gene is selected from the group consisting of TOM70, TIM22, TOM40, Imp2, mitochondrial Hsp70, ATM1-ABC transporter proteins, Frataxin, Ferredoxin, ERV1, ferredoxin, NADPH oxido-reductase [FNR], pyruvate dehydrogenase α subunit, pyruvate dehydrogenase β subunit, mitochondrial glycerol-3-phosphate dehydrogenase (mtG3PDH), manganese-containing superoxide dismutase (MnSOD), DNAJ (Hsp70 interacting), Iron Sulfur cluster ISU1, Cystein desulfurase Nsfl, NAR1, RLI1, ATPase subunit A, RNA polymerase I, RNA polymerase III, Inhibitor of apoptosis (IAP), and FAS apoptotic. 
     
     
         22 . The method of  claim 20 , wherein said one or more bacteria express an inhibitor of a pathogen or a parasite specific gene product, thereby reducing the susceptibility of the bee to a disease or disorder selected from the group consisting of Colony Collapse Disorder (CCD) and infection. 
     
     
         23 . The method of  claim 22 , wherein the pathogen or parasite is selected from the group consisting of Acute Bee Paralysis Virus (ABPV), Kashmir Bee Virus (KBV), Israeli Acute Paralysis Virus (IAPV),  Nosema ceranae, Nosema cerana, Nosema apis , Deformed Wing Virus, and  Varroa destructor  mite. 
     
     
         24 . The method of  claim 22 , wherein the bacteria express at least two noncontiguous dsRNAs downregulating expression of a pathogen or parasite specific gene product. 
     
     
         25 . A method for reducing the susceptibility of a bee colony to infestation by the Small Hive Beetle, comprising administering an effective amount of a composition comprising one or more bacteria genetically engineered to express at least one heterologous nucleic acid, wherein one or more bacteria are native to the microbiome of a host bee to hive components, wherein said bacteria express an inhibitor of a Small Hive Beetle specific gene product. 
     
     
         26 . The method of  claim 25 , wherein the bacteria express at least two noncontiguous dsRNAs downregulating expression of a Small Hive Beetle specific gene product. 
     
     
         27 . A method for expression of a heterologous nucleic acid sequence in a bee, the method comprising administering to the bee at least one modified  Snodgrassella alvi  bacterium comprising an expression plasmid for expression of the heterologous nucleic acid sequence. 
     
     
         28 . The method of  claim 27 , wherein the heterologous nucleic acid sequence encodes a molecule selected from the group consisting of a protein, a peptide, an antisense DNA, an inhibitory RNA, a dsRNA, a siRNA, a shRNA, a sgRNA and a miRNA.