miR-302 family, miR-145 and miR-296 temporal expression profile along the cell cycle of human pluripotent stem cells

Rodríguez-Varela, María Soledad; Mucci, Sofía; Videla-Richardson, Guillermo A.; Isaja, Luciana; Sevlever, Gustavo; Scassa, María Élida; Romorini, Leonardo

doi:10.1016/j.gep.2021.119168

Cited by 2 publications

(4 citation statements)

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“…500 ng of total RNA was used for cDNA synthesis with 15 mM of random hexamers and MMLV reverse transcriptase (Promega), according to the manufacturer’s instructions. For miRNA reverse transcription, cDNA was generated using SuperScript™ II Reverse Transcriptase (Thermo Scientific) and miRNA-specific stem-loop primers as previously described [14]. For real-time PCR, cDNA samples were diluted 5-folds and PCR was performed with StepOne Plus Real-Time PCR System (PE Applied Biosystems).…”

Section: Methodsmentioning

confidence: 99%

“…Interestingly, some miRNAs promote hPSC proliferation [13], and some are regulated in a cell cycle-dependent manner [14].…”

Section: Introductionmentioning

confidence: 99%

“…Particularly, microRNAs (miRNAs), which are non-coding RNA molecules of approximately 18-24 nucleotides that regulate gene expression by interacting with specific sequences present in mRNAs, are implicated in fine-tuning numerous biological processes such as cell cycle progression, proliferation, and differentiation [12]. Interestingly, some miRNAs promote hPSC proliferation [13], and some are regulated in a cell cycle-dependent manner [14].…”

Section: Introductionmentioning

confidence: 99%

“…The copyright holder for this preprint this version posted March 2, 2024. ; https://doi.org/10.1101/2024.02. 28.582539 doi: bioRxiv preprint Interestingly, some miRNAs promote hPSC proliferation [13], and some are regulated in a cell cycle-dependent manner [14].…”

Section: Introductionmentioning

confidence: 99%

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“Identification of microRNAs regulated by E2F transcription factors in human pluripotent stem cells”

Rodríguez-Varela,

Vautier,

Mucci

et al. 2024

Preprint

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Human pluripotent stem cells (hPSCs), which include embryonic and induced pluripotent stem cells (hESCs and hiPSCs, respectively), have an unusual cell cycle structure which consists of a short G1 phase and the absence of the G1/S checkpoint regulation. E2F transcription factors (E2Fs) play an important role in the G1/S transition. G1 duration contributes to hPSC fate determination, and microRNAs (miRNAs) play critical roles in this commitment. As little is known about the interplay between E2Fs and miRNAs in hPSCs, we aimed to identify miRNAs that are regulated by E2Fs in these cells. We first found that mRNA expression levels of canonical E2F repressors were more expressed than most E2F activators in G1-arrested hPSCs. Moreover, we observed higher mRNA and protein expression levels of canonicalE2F2,E2F3A,andE2F5in G1 synchronized hPSCs compared to human fibroblasts (HF). However,E2F1andE2F4protein expression levels were higher in HF. We next found that E2F inhibition with HLM006474 induced an increase in the G1 cell population without affecting hPSC viability, concomitantly with a decrease inOCT-4mRNA levels and the percentage of OCT-4+hPSCs. Next, by RNA-seq analysis we found 52 differentially expressed (DEGs) miRNAs in HLM006474-treated hESCs. RT-qPCR validation of some of the DEGs let us conclude that miR-19a-3p, miR-19b-3p, miR-4454, miR-1260a, miR-1260b, miR-454-3p and miR-301a-3p are regulated by E2Fs in hPSCs. Interestingly, gene target and ontology analysis of these miRNAs revealed a possible implication in proliferation and cell cycle regulation, development, and neural differentiation.

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Section: Methodsmentioning

confidence: 99%

“…Interestingly, some miRNAs promote hPSC proliferation [13], and some are regulated in a cell cycle-dependent manner [14].…”

Section: Introductionmentioning

confidence: 99%