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Profiling of RNA-binding protein LSM14B target sites in oocyte by LACE-seq

ABSTRACT: Profiling of RNA-binding protein LSM14B target sites in oocyte by LACE-seq

PROVIDER: PRJNA849588 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR19663574.fastq.gz	Fastqsanger.gz
	SRR19663575.fastq.gz	Fastqsanger.gz

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Profiling of RNA-binding protein LSM14B target sites in oocyte by LACE-seq

Project description:RNA-binding proteins (RBPs) have essential functions during oocyte development. In order to explore the regulatory role of RNA-binding protein LSM14B in oocyte, we determined the binding sites and regulatory mechanisms for LSM14B by LACE-seq.

2023-09-08 | GSE206190 | GEO

LSM14B is essential for oocyte meiotic maturation by regulating maternal mRNA storage and clearance

Project description:Fully grown oocytes remain transcriptionally quiescent, yet many maternal mRNAs are synthesized and retained in growing oocytes. We now know that maternal mRNAs are stored in a structure called the mitochondria associated ribonucleoprotein domain (MARDO). But the components and functions of MARDO remain elusive. Here, we found that LSM14B knockout prevents the proper storage and timely clearance of mRNAs (including Cyclin B1, Btg4, and other mRNAs that are translationally activated during meiotic maturation), specifically by disrupting MARDO assembly during oocyte growth and meiotic maturation. With decreased levels of storage and clearance, the LSM14B knockout oocytes failed to enter meiosis II, ultimately resulting in female infertility. Our results demonstrate the function of LSM14B in MARDO assembly, couple the MARDO with mRNA clearance and oocyte meiotic maturation.

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Global Profiling of RNA-Binding Protein Target Sites by LACE-seq

Project description:Global Profiling of RNA-Binding Protein Target Sites by LACE-seq

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Global Profiling of RNA-Binding Protein Target Sites by LACE-seq

Project description:RNA-binding proteins (RBPs) have essential functions during germline and early embryo development. However, current methods are unable to identify the in vivo targets of an RBP in those rare cells. Here, by coupling RBP-mediated reverse transcription termination with linear amplification of cDNA ends and sequencing, we present the LACE-seq method for identifying RBP-regulated RNA networks at or near the single-oocyte level. We determined the binding sites and regulatory mechanisms for several RBPs, including Argonaute 2 (Ago2), Mili, Ddx4, and PTBP1, in mature oocytes. Unexpectedly, transcriptomic and proteomic analysis of Ago2-/- oocytes revealed that Ago2 interacted with endogenous small interfering RNAs (endo-siRNAs) to repress mRNA translation globally. Furthermore, the Ago2 and endo-siRNA complexes could fine-tune the transcriptome by slicing long terminal repeat retrotransposon-derived chimeric transcripts. The precise mapping of RBP binding sites in a few cells opens the door for studying the roles of RBPs in embryonic development and reproductive diseases.

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LSM14B is essential for oocyte meiotic maturation by regulating maternal mRNA storage and clearance

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Identifying the Translatome of WT and Lsm14b KO GV-Stage Oocytes in mice via Scarce Sample Polysome Profiling (SSP-profiling)

Project description:RNA-binding proteins (RBPs) have essential functions during oocyte development.In order to explore the regulatory role of RNA-binding protein LSM14B in oocyte, we Identified the translatome of WT and Lsm14b KO GV-Stage Oocytes in mice via Scarce Sample Polysome Profiling (SSP-profiling).

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Comparing the Transcriptomes of WT oocytes with Lsm14b KO oocytes by RNA-Seq Analysis

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LSM14B is essential for oocyte meiotic maturation by regulating maternal mRNA storage and clearance

Project description:LSM14B is essential for oocyte meiotic maturation by regulating maternal mRNA storage and clearance

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Loss of LSM14b affects the meiosis process of mouse oocytes

Project description:RNA-Seq can comprehensively and quickly obtain almost all transcripts and information of specific cells in rustic states.Here,we compared up-regulated genes in GV and MII oocytes from LSM14b deficient mice and wild-type mice by RNA-Seq,highlighting the important role of LSM14b in meiosis of mouse oocytes.

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