Induction of hemogenic endothelium from pluripotent stem cells
Described herein are methods and related compositions for inducing differentiation of human pluripotent stem cells (hPSCs) into hemogenic endothelium with pan-myeloid potential or restricted potential, by forced expression in the hPSCs of a combination of transcription factors as described herein.
1. A kit for hemogenic reprogramming, comprising:
one or more recombinant expression viruses comprising a nucleic acid encoding an ETV2 or ERG protein, and a GATA1 protein.
2. The kit of claim 1 , wherein the ETV2, ERG, or GATA1 sequence are from human, mouse, or rat.
3. A kit for hemogenic reprogramming, comprising:
one or more isolated modified mRNAs comprising an open reading frame for ETV2 or ERG, and GATA1, wherein the modified mRNA comprises:
a 5′ synthetic cap;
modified nucleotides, wherein the modified nucleotides comprise a member selected form the group consisting of pseudo-uridine and 5-methyl-cytosine; and
a 3′ poly-A tail.
4. The kit of claim 3 , wherein the 5′ synthetic cap is methylated.
5. A kit for hemogenic reprogramming, comprising:
one or more isolated modified mRNAs comprising an open reading frame from ETV2 or ERG, and GATA1, wherein the modified mRNA comprises:
a 5′ synthetic cap;
modified nucleotides; and
a poly-A tail, wherein the modified mRNA comprises 3′-O-Me-m7G(5′)pp(5′)G.
6. The kit of claim 1 , wherein the ETV2 protein comprises SEQ ID NO:1.
7. The kit of claim 1 , wherein the ERG protein comprises SEQ ID NO:3.
8. The kit of claim 1 , wherein the GATA protein comprises SEQ ID NO:2.
9. A kit for hemogenic reprogramming, comprising:
one or more double stranded DNA expression vectors comprising a heterologous nucleic acid sequence encoding an ETV2 or ERG protein and a GATA1 protein.
10. The kit of claim 9 , wherein the ETV2, ERG, or GATA1 sequence are from human, mouse, or rat.
11. The kit of claim 9 , wherein the ETV2 protein comprises SEQ ID NO:1.
12. The kit of claim 9 , wherein the ERG protein comprises SEQ ID NO:3.
13. The kit of claim 9 , wherein the GATA protein comprises SEQ ID NO:2.
14. The kit of claim 1 , wherein the ETV2, ERG, or GATA1 sequence are from human, mouse, or rat.
15. The kit of claim 3 , wherein the ETV2 protein comprises SEQ ID NO:1.
16. The kit of claim 3 , wherein the ERG protein comprises SEQ ID NO:3.
17. The kit of claim 3 , wherein the GATA protein comprises SEQ ID NO:2.
18. The kit of claim 5 , wherein the ETV2, ERG, or GATA1 sequence are from human, mouse, or rat.
19. The kit of claim 5 , wherein the ETV2 protein comprises SEQ ID NO:1.
20. The kit of claim 5 , wherein the ERG protein comprises SEQ ID NO:3.
21. The kit of claim 5 , wherein the GATA protein comprises SEQ ID NO:2.