Methods and compositions for increased double stranded RNA production
AssigneeApse, Inc.
InventorsJohn L. Killmer, Patrick D. McLaughlin, Juan Pedro Humberto Arhancet
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Abstract
Methods and materials for improved in vivo production of dsRNA are presented. Yields of dsRNA are significantly increased in the presence of capsid protein. The improved yield of dsRNA is not dependent on the presence of specific cognate binding sites for capsid protein associated with the dsRNA, but is dependent on capsid protein.
Claims33
A method for producing recoverable unencapsidated dsRNA in a microbial cell, the microbial cell comprising (i) a leviviridae coat protein gene encoding a capsid protein selected from MS2, Qβ and an amino-terminal fragment of MS2 or Qβ and (ii) an expression cassette comprising a promoter operably linked to a DNA sequence encoding a heterologous dsRNA molecule, said dsRNA molecule comprising a self-complementary stretch of sequence separated by non-complementary sequence such that upon hybridization of the complementary sequences a stem-loop structure is formed, the stem structure having a length exceeding the interior diameter of an MS2 capsid, the method comprising the step of culturing the microbial cell under conditions such that the dsRNA and the coat protein gene are co-expressed in the microbial cell, wherein the amount of unencapsidated dsRNA produced in the microbial cell is significantly increased compared to the amount of unencapsidated dsRNA produced in the microbial cell when the coat protein gene is not co-expressed.
Dependencies are inferred from the claim text.