Method for producing self-assembling paramyxoviral nucleocapsid-like particles and their uses
AssigneesCentre National De La Recherche Scientifique, Universite Grenoble Alpes, Commissariat A L'energie Atomique Et Aux Energies Alternatives
InventorsMartin Blackledge, Marlène Ringkjobing-Jensen, Sigrid Milles, Robertus Ruigrok, Guy Schoehn, Guillaume Communie
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Abstract
Embodiments of the disclosure relate to the domain of virology of Paramyxoviruses. The disclosure concerns a method for producing self-assembling paramyxoviral particles and a method for identifying a compound able to inhibit the replication or the transcription of a Paramyxovirus. The disclosure also pertains to the nucleocapsid-like particles obtainable by the method of the invention and their use for biotechnological and pharmaceutical applications.
Claims7
A method for producing self-assembling paramyxoviral nucleocapsid-like particles comprising the steps of:
- a. co-expressing recombinant nucleoprotein (N) and phosphoprotein (P) proteins in order to allow the formation of NºP complexes wherein:
- i. the recombinant N protein comprises an NCORE domain including a C-terminal domain (CTD) arm and an N-terminal domain (NTD) arm, and
- ii. the recombinant P protein comprises an N-binding domain;
- b. purifying NºP complexes, the purified NºP complexes comprising recombinant N and P proteins only and being RNA-free;
- c. adding RNA molecules of interest to the purified NºP complexes, wherein the RNA molecules of interest comprise at least 6 nucleotides and are not poly-U homopolymers;
- and
- d. recovering the resulting nucleocapsid-like particles.
A method for identifying a compound able to inhibit the replication or transcription of a Paramyxovirus comprising the steps of:
- a. co-expressing recombinant N and P proteins in order to allow the formation of NºP complex wherein:
- i. the recombinant N protein comprises an N-core domain including a CTD arm and an NTD arm, and
- ii. the recombinant P protein comprises an N-binding domain;
- b. purifying NºP complexes, the purified NºP complex comprising recombinant N and P proteins only and being RNA-free;
- c. adding a compound to be tested;
- d. adding a RNA molecule wherein said RNA molecule comprises at least 6 nucleotides and is not a poly-U homopolymer;
- e. detecting the presence of nucleocapsid-like particles in comparison with a control wherein no compound is present;
- and
- f. identifying a compound able to inhibit the replication or transcription of a Paramyxovirus, where the assembly of nucleocapsid-like particles is inhibited in the presence of such compound, compared to control.
Dependencies are inferred from the claim text.