A CRISPR RNA-binding protein screen reveals regulators of RUNX1 isoform generation.

Autor:	Davis AG; Moores Cancer Center.; Department of Biological Sciences., Einstein JM; Department of Cellular and Molecular Medicine.; Stem Cell Program, and.; Institute for Genomic Medicine, University of California, San Diego, La Jolla, CA., Zheng D; Department of Microbiology, Biochemistry, and Molecular Genetics, Rutgers New Jersey Medical School, Newark, NJ., Jayne ND; Moores Cancer Center.; Department of Biological Sciences., Fu XD; Department of Cellular and Molecular Medicine., Tian B; Department of Microbiology, Biochemistry, and Molecular Genetics, Rutgers New Jersey Medical School, Newark, NJ.; Program in Gene Expression and Regulation, Center for Systems and Computational Biology, Wistar Institute, Philadelphia, PA; and., Yeo GW; Department of Cellular and Molecular Medicine.; Stem Cell Program, and.; Institute for Genomic Medicine, University of California, San Diego, La Jolla, CA., Zhang DE; Moores Cancer Center.; Department of Biological Sciences.; Department of Pathology, University of California, San Diego, La Jolla, CA.
Jazyk:	angličtina
Zdroj:	Blood advances [Blood Adv] 2021 Mar 09; Vol. 5 (5), pp. 1310-1323.
DOI:	10.1182/bloodadvances.2020002090
Abstrakt:	The proper balance of hematopoietic stem cell (HSC) self-renewal and differentiation is critical for normal hematopoiesis and is disrupted in hematologic malignancy. Among regulators of HSC fate, transcription factors have a well-defined central role, and mutations promote malignant transformation. More recently, studies have illuminated the importance of posttranscriptional regulation by RNA-binding proteins (RBPs) in hematopoiesis and leukemia development. However, the RBPs involved and the breadth of regulation are only beginning to be elucidated. Furthermore, the intersection between posttranscriptional regulation and hematopoietic transcription factor function is poorly understood. Here, we studied the posttranscriptional regulation of RUNX1, a key hematopoietic transcription factor. Alternative polyadenylation (APA) of RUNX1 produces functionally antagonistic protein isoforms (RUNX1a vs RUNX1b/c) that mediate HSC self-renewal vs differentiation, an RNA-processing event that is dysregulated in malignancy. Consequently, RBPs that regulate this event directly contribute to healthy and aberrant hematopoiesis. We modeled RUNX1 APA using a split GFP minigene reporter and confirmed the sensitivity of our model to detect changes in RNA processing. We used this reporter in a clustered regularly interspaced short palindromic repeats (CRISPR) screen consisting of single guide RNAs exclusively targeting RBPs and uncovered HNRNPA1 and KHDRBS1 as antagonistic regulators of RUNX1a isoform generation. Overall, our study provides mechanistic insight into the posttranscriptional regulation of a key hematopoietic transcription factor and identifies RBPs that may have widespread and important functions in hematopoiesis. (© 2021 by The American Society of Hematology.)
Databáze:	MEDLINE
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