Virus-like particles and use thereof
A virus-like particle encapsulating a target protein is provided. The virus-like particle contain a Gag protein, and the Gag protein forms a dimer with the target protein.
1. A virus-like particle encapsulating a Cas family protein and a gRNA, comprising:
a fusion protein of FKBP12 and Gag protein and a fusion protein of FRB and the Cas family protein, or a fusion protein of FRB and Gag protein and a fusion protein of FKBP12 and a Cas family protein;
an mRNA or a self-cleavage product of the mRNA wherein the mRNA having the gRNA sequence interposed between a first ribozyme sequence and a second ribozyme sequence and a packaging signal sequence; and
a rapamycin or a rapamycin derivative;
wherein the FKBP12, the rapamycin or the rapamycin derivative, and the FRB are bound together and the Gag protein forms a dimer with the Cas family protein.
2. A method for manufacturing a genome-edited cell, comprising:
inoculating the cell with the virus-like particle according to claim 1 .
3. A kit for manufacturing the virus-like particle according to claim 1 , comprising:
an expression vector for a fusion protein of FKBP12 and Gag protein and an expression vector for a fusion protein of FRB and the Cas family protein; or an expression vector for a fusion protein of FRB and Gag protein and an expression vector for a fusion protein of FKBP12 and the Cas family protein; and
an expression vector for mRNA having a base sequence of a target RNA or a multiple cloning site, which is interposed between a first ribozyme sequence and a second ribozyme sequence, and a packaging signal sequence.
4. A method for treating diseases caused by genetic mutation, infections, or cancer, comprising:
administering an effective dose of the virus-like particle according to claim 1 to a patient in need of treatment.
5. A method for manufacturing a virus-like particle encapsulating a Cas family protein and a gRNA, comprising the following (1) and (2):
(1) letting a cell to express:
a fusion protein of FKBP12 and Gag protein and a fusion protein of FRB and the Cas family protein, or a fusion protein of FRB and Gag protein and a fusion protein of FKBP12 and a Cas family protein, and
an mRNA having the gRNA sequence interposed between a first ribozyme sequence and a second ribozyme sequence and a packaging signal sequence in the presence of rapamycin or a rapamycin derivative; and
(2) obtaining a medium containing the virus-like particle encapsulating the Cas family protein and the gRNA.
6. The method for manufacturing according to claim 5 ,
wherein in (1), a nucleic acid encoding the fusion protein or the mRNA is introduced into the cell by lipofection or electroporation.