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Genome-Scale CRISPRa Screen Identifies Novel Factors for Cellular Reprogramming.
Jian Yang1, Sandeep S Rajan2, Mathias J Friedrich1
1Wellcome Trust Sanger Institute, Genome Campus, Hinxton, Cambridgeshire CB10 1SA, UK.
Stem Cell Reports
|March 26, 2019
Summary
Scientists used CRISPR activation to discover new genes that help revert cells to a pluripotent state. Sall1 was identified as a key factor that enhances reprogramming and maintains pluripotency, offering new insights into cellular reprogramming mechanisms.
Area of Science:
- Cell Biology
- Genetics
- Developmental Biology
Background:
- Epiblast stem cells (EpiSCs) can be reverted to a pluripotent embryonic stem cell (ESC)-like state.
- Understanding the genetic factors that drive this cellular reprogramming is crucial for regenerative medicine.
Purpose of the Study:
- To perform a genome-scale screen to identify novel genes involved in cellular reprogramming.
- To investigate the role of Sall1 in enhancing the reprogramming of mouse EpiSCs and fibroblasts.
Main Methods:
- CRISPR activation screening in EpiSCs to identify candidate reprogramming factors.
- Validation of candidate genes, including Sall1, for their ability to induce pluripotency.
- RNA sequencing to identify downstream targets of Sall1 and Nanog.
Main Results:
- Identified 142 candidate genes, validating 50 previously unknown reprogramming factors.
- Sall1 significantly augments reprogramming of mouse EpiSCs and fibroblasts, producing fully pluripotent cells.
- Sall1 synergizes with Nanog, delays ESC to EpiSC conversion, and has Klf5 and Fam189a2 as downstream targets.
Conclusions:
- CRISPR activation is a powerful tool for dissecting complex cellular processes like reprogramming.
- Sall1 is a novel and potent factor that enhances cellular reprogramming and maintains pluripotency.
- Identified new downstream targets of Sall1 and Nanog, advancing the understanding of reprogramming pathways.
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