Method for Producing Induced Pluripotent Stem Cells
AssigneeNew England Biolabs, Inc.
InventorsDapeng Sun, Fanfan Chen, Xianghui Li, Ling Yu, Yanye Feng
Life of this patent
Estimating the term…
Abstract
Described herein is an inactivated viral particle comprising one or more transcription factor proteins packaged within the particle. A method for using the particle to make induced pluripotent stem cells is also provided.
Claims32
An inactivated viral particle comprising:
- (a) an envelope;
- and
- (b) one or more isolated transcription factor proteins.
An inactivated viral particle wherein the one or more transcription factor proteins are selected from the group consisting of Sox2, Oct4, Klf4, c-Myc, C/EBPβ, GATA3 and NeuroD1, packaged within the particle.
The method of claim 20, further comprising transplanting the recellularized organ or tissue into a recipient subject.
A screening method comprising:
- (a) transfecting somatic cells with an inactivated viral particle wherein the inactivated virus particle comprises:
- (i) an envelope;
- and one or more isolated transcription factor proteins, or
- (ii) an envelope and one or more transcription factor proteins selected from the group consisting of Sox2, Oct4, Klf4, c-Myc, C/EBPβ, GATA3 and NeuroD1, packaged within the particle;
- or
- (iii) an inactivated Sendai virus, herpesvirus, parainfluenza virus or lentivirus particle comprising an HVJ envelope and one or more of the isolated transcription factor proteins packaged within the particle;
- or
- (iv) an inactivated Sendai viral particle comprising an HVJ envelope;
- or
- (v) a Sox2 transcription factor having an amino acid sequence that is at least 80% identical to a mammalian Sox2 protein;
- an Oct4 transcription factor, having an amino acid sequence that is at least 80% identical to mammalian Oct4 protein;
- a Klf4 transcription factor, having an amino acid sequence that is at least 80% identical to mammalian Klf4 protein;
- and c-Myc transcription factor, having an amino acid sequence that is at least 80% identical to mammalian c-Myc protein;
- packaged within the particle;
- or
- (vi) an Oct4 transcription factor, having an amino acid sequence that is at least 80% identical to a mammalian Oct4 protein;
- and an C/EBPβ transcription factor, having an amino acid sequence that is at least 80% identical to mammalian C/EBPβ protein;
- packaged within the particle;
- or
- (vii) a Sox2 transcription factor having an amino acid sequence that is at least 80% identical to a mammalian Sox2 protein;
- an GATA3 transcription factor having an amino acid sequence that is at least 80% identical to mammalian GATA3 protein;
- and
- (b) contacting a test agent with the somatic cells;
- (c) culturing the somatic cells;
- and
- (d) determining whether the test agent has any effect on the cell type produced by culturing step (c).
A screening method comprising:
- (a) packaging a test agent within an inactivated viral particle in the absence of isolated transcription factor proteins or nucleic acid encoding the same;
- (b) transfecting an induced pluripotent stem cell with the inactivated viral particle of step (a);
- (c) culturing the transfected cells on a differentiation medium;
- and
- (d) determining whether the test agent has any effect on the cell type produced by culturing step.
Dependencies are inferred from the claim text.