Compositions and methods for increasing the efficiency of cell cultures used for food production
Provided herein are compositions and methods to make and use engineered cells, for the purpose of increasing the cell density of a culture comprising metazoan cells and for the production of a cultured edible product.
1. A method for increasing the cell density of a culture comprising metazoan cells having myogenic or fibroblastic capacity, the method comprising:
a) introducing into the cells one or more polynucleotide sequences encoding glutamine synthetase (GS), insulin-like growth factor (IGF), albumin or a combination thereof,
b) introducing into the cells a polynucleotide sequence encoding a telomerase reverse transcriptase (TERT), and
c) culturing the cells in a cultivation infrastructure.
2. The method of claim 1 , wherein the cells comprise a loss-of-function mutation in one or more genes encoding cyclin-dependent kinase inhibitor (CKI) proteins.
3. The method of claim 2 , wherein the CKI proteins are p15, p16, paralogs, orthologs, or genetic variants thereof.
4. The method of claim 1 , wherein the cells are from a self-renewing cell line.
5. The method of claim 1 , wherein the cells are further modified to express a myogenic transcription factor.
6. The method of claim 1 , wherein:
a) when the polynucleotide encoding GS is introduced into the cells, the concentration of glutamine in the culture medium is increased when compared to cultures of cells in which the polynucleotide encoding GS is not introduced into the cells;
b) when the polynucleotide encoding IGF is introduced into the cells, the concentration of IGF in the culture medium is increased when compared to cultures of cells in which the polynucleotide encoding IGF is not introduced into the cells; and/or
c) when the polynucleotide encoding albumin is introduced into the cells, the concentration of albumin in the culture medium is increased when compared to cultures of cells in which the polynucleotide encoding albumin is not introduced into the cells.
7. The method of claim 1 , wherein the cells are further modified to inhibit the HIPPO signaling pathway.
8. The method of claim 7 , wherein inhibiting the HIPPO signaling pathway comprises activating Yes-Associated Protein 1 (YAP1), Transcriptional co-Activator with PDZ-binding motif (TAZ), or a combination thereof in the cells.
9. The method of claim 1 , wherein the cells are from a livestock, poultry, game or aquatic animal species.
10. The method of claim 1 , wherein the cells are from a chicken, duck, or turkey.
11. The method of claim 1 , wherein the cells are from a fish.
12. The method of claim 1 , wherein the cells are from a livestock species.
13. The method of claim 12 , wherein the livestock species is porcine or bovine.
14. The method of claim 1 , wherein the cells are from any animal species intended for human or non-human dietary consumption.
15. The method of claim 1 , wherein the cells are myogenic cells.
16. The method of claim 1 , wherein the cells are non-myogenic cells modified to become myogenic cells through expression of one or more myogenic transcription factors.
17. The method of claim 1 , wherein the polynucleotide sequence encoding GS comprises a GS coding sequence selected from SEQ ID NOs: 18, 21, and 24, wherein the polynucleotide sequence encoding IGF comprises an IGF coding sequence selected from SEQ ID NOs: 1-6, 15, 16, 19, 20, 23 and 25, or wherein the polynucleotide sequence encoding albumin comprises an albumin coding sequence selected from SEQ ID NOs: 7-11, 17, and 22.
18. The method of claim 1 , wherein the cells further have the capacity for skeletal muscle tissue specification.