|
| Status |
Public on Jul 17, 2015 |
| Title |
Integrative analyses of human reprogramming reveal dynamic nature of induced pluripotency [shRNA] |
| Organism |
Homo sapiens |
| Experiment type |
Other
|
| Summary |
Induced pluripotency is a promising avenue for disease modeling and therapy, but the molecular principles underlying this process, particularly in human cells, remain poorly understood due to donor-to-donor variability and intercellular heterogeneity. Here we constructed and characterized a clonal, inducible human reprogramming system that provides a reliable source of cells at any stage of the process. This system enabled integrative transcriptional and epigenomic analysis across the human reprogramming timeline at high resolution. We observed distinct waves of gene network activation, including the ordered re-activation of broad developmental regulators followed by early embryonic patterning genes and culminating in the emergence of a signature reminiscent of pre-implantation stages. Moreover, complementary functional analyses allowed us to identify and validate novel regulators of the reprogramming process. Altogether, this study sheds light on the molecular underpinnings of induced pluripotency in human cells and provides a robust cell platform for further studies.
|
| |
|
| Overall design |
We infected hiF-T cells in biological replicate with a shRNA library (2,900 shRNA targeting 370 distinct epigenetic regulators). This library is a sub-pool of the human 45K shRNA pool (Luo et al., 2008) distributed by the Broad Institute Genomic Perturbations Platform and the RNAi Consortium (TRC). The plasmid for shRNA expression under the control of the constitutive U6 snRNA promoter was the lentiviral vector pLKO.1. shRNA pool production and infection conditions were performed as previously described (Luo et al., 2008). 24h hours after infection as well as at 15 days of repgrogramming, we extracted the genomic DNA and detrmined the abundance of distinct shRNAs based on their representation in the genomic DNA.
|
| |
|
| Contributor(s) |
Cacchiarelli D, Trapnell C, J. Ziller M, Ho Sui SJ |
| Citation(s) |
26186193 |
| |
| Submission date |
Jul 17, 2015 |
| Last update date |
May 15, 2019 |
| Contact name |
Shannan J Ho Sui |
| E-mail(s) |
shosui@hsph.harvard.edu
|
| Organization name |
Harvard T.H. han Scholol of Public Health
|
| Department |
Biostatistics
|
| Street address |
655 Huntington Ave, Building 2, Rm 437A
|
| City |
Boston |
| State/province |
Massachusetts |
| ZIP/Postal code |
02115 |
| Country |
USA |
| |
|
| Platforms (1) |
|
| Samples (4)
|
|
| This SubSeries is part of SuperSeries: |
| GSE62777 |
Integrative analyses of human reprogramming reveal dynamic nature of induced pluripotency |
|
| Relations |
| BioProject |
PRJNA290154 |
| SRA |
SRP061265 |
Less...
More...