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US 2025/0051774 A1Published application
DIRECT PRODUCTION OF SIRNAS IN SACCHAROMYCES BOULARDII AND PACKAGING IN EXTRACELLULAR VESICLES (EVs) FOR TARGETED GENE SILENCING
AssigneeMercury Bio, Inc.
InventorsTatiana Vinogradova-Shah, Richard T. Sayre, Alex Pertzev, Nianzi Liu, Arle Kruckeberg
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Abstract
The present invention is direct to the direct expression of siRNA molecules in yeast, which can be packaged into extracellular vesicles (EVs) and delivered to a target organism causing the downregulation of a select gene.
Claims67
An extracellular vesicle composition comprising:
- a quantity of isolated yeast-generated extracellular vesicle (EVs) containing one or more heterologous small interfering RNAs (siRNAs);
- and
- wherein said heterologous siRNAs were produced from a double-stranded RNA (dsRNA) heterologously expressed in the Saccharomyces cell, and further processed into said siRNAs by a heterologously expressed fusion peptide having a first dicer protein domain fused to an second EV membrane protein domain.
A system for the production of extracellular vesicles containing heterologous RNA molecules comprising:
- a yeast cell expressing heterologous nucleotide, operably linked to a promoter, encoding:
- one or more heterologous double-stranded RNA (dsRNA);
- fusion peptide having a first dicer protein domain fused to an second EV membrane protein domain;
- and
- wherein fusion peptide is anchored in an extracellular vesicle (EVs) of the cell, and further processes the dsRNA into said small interfering RNAs (siRNAs).
2 more elements
A method of producing extracellular vesicles containing siRNAs, the method comprising:
- transforming a yeast host cell to express a heterologous nucleotide, operably linked to a promoter, encoding:
- one or more heterologous double-stranded RNA (dsRNA);
- a heterologous EV membrane protein linked to a dicer enzyme that processes said dsRNAs into siRNAs;
- a dsRNA directed to inhibit the expression of more target genes in a host cell;
- culturing the host cell under conditions that promote extracellular vesicle (EV) generation and
- isolating the EVs from the culture, wherein said isolated EVs contain the one or more heterologous siRNAs.
3 more elements
Dependencies are inferred from the claim text.