Methods and compositions for bat ipsc preparation and use
AssigneeIcahn School of Medicine at Mount Sinai
InventorsThomas P. Zwaka, Marion Dejosez Zwaka, Adolfo Garcia- Sastre
Life of this patent
Estimating the term…
Abstract
Disclosed herein are compositions and methods of making and using bat IPSCs (BipS). Also disclosed herein are methods and compositions of virus nucleic acids residing in bat IPSCs. Also disclosed are nucleotides, cells, and methods associated with the compositions including their use as vaccines.
Claims62
An induced pluripotent bat stem cell (bat IPSC), wherein the cell is in a pluripotent state.
A method of producing induced pluripotent bat stem cells (bat IPSCs), the method comprising:
- (i) reprogramming isolated bat cells with Oct4, Sox2, cMyc, and Klf4 factors;
- (ii) culturing the reprogrammed cells on feeder cells in a medium comprising FGF, Leukemia inhibitory factor (Lif), SCF, and Forskolin until colonies appear;
- and
- (iii) splitting cells using a low concentration EDTA buffer;
- thereby producing IPSCs from bats.
A method of producing induced pluripotent bat stem cells (bat IPSCs), the method comprising:
- (i) reprogramming isolated bat cells with Oct4, Sox2, cMyc, and Klf4 factors;
- (ii) culturing the reprogrammed cells in feeder free medium comprising FGF, Leukemia inhibitory factor (Lif), SCF, and Forskolin until colonies appear;
- and
- (iii) splitting cells using a low concentration EDTA buffer
- thereby producing IPSCs from bats.
A composition for reprogramming a bat cell to produce pluripotent stem cells comprising a medium comprising FGF, Leukemia inhibitory factor (Lif), SCF, and Forskolin.
A method of obtaining viral sequences from bat IPSCs, the method comprising
- obtaining bat IPSCs;
- identifying viral sequences residing in the bat iPSC genome or intracellular virus genome;
- and
- assembling the viral sequences;
- thereby obtaining viral sequences from the bat iPSCs.
A method of obtaining viral sequences from virus particles shed by bat IPSCs or cells derived from bat IPSCs, the method comprising
- obtaining bat IPSCs or cells derived from bat IPSCs;
- culturing the bat IPSCs or cells derived from bat IPSCs under conditions that allows shedding of virus particles into the culture media;
- collecting the culture media;
- identifying viral sequences residing in the culture media;
- and
- assembling the viral sequences,
- thereby obtaining viral sequences from virus particles shed by bat iPSCs or cells derived from bat IPSCs.
A recombinant nucleic acid molecule, comprising
- a promoter, and
- a nucleic acid selected from SEQ ID NO:
- 1-349 encoding for a viral protein or fragment thereof.
An expression vector comprising
- a promoter and
- a nucleic acid selected from SEQ ID NO:
- 1-349 encoding for a viral protein or fragment thereof.
An isolated protein or peptide comprising an amino acid sequence encoded in a nucleic acid set forth in SEQ ID NO:
- 1-349, wherein the peptide is no more than 100 amino acids in length, and an optional pharmaceutically acceptable carrier.
A recombinant cell comprising a nucleic acid or a portion of a nucleic acid set forth in SEQ ID NO:
- 1-349.
A recombinant cell comprising a protein or a portion of a protein encoded by a nucleic acid set forth in SEQ ID NO:
- 1-349.
A composition comprising an inhibitor of a protein encoded by a nucleic acid selected from SEQ ID NO:
- 1-349.
Dependencies are inferred from the claim text.