a-uv-based-chirp-method-for-verifiable-identification-proteins-that

A UV-Based ChIRP Method For The Verifiable Identification of Proteins That Directly Interact With LncRNA: From Cultured Cells To Mouse Testis

Long non-coding RNAs (lncRNAs) have emerged as important regulators of diverse biological processes. However, the molecular mechanisms underlying the ...

Nanjing Medical University

Shanghai Jiao Tong University

Long non-coding RNAs (lncRNAs) have been the focus of ongoing research in a diversity of cellular processes. LncRNAs are abundant in mammalian testis, but their biological function remains poorly known.

Microinjection of antisense oligonucleotides into living mouse testis enables lncRNA function study.

Controversy continues over the functional prevalence of long non-coding RNAs (lncRNAs) despite their being widely investigated in all kinds of cells and organisms. In animals, lncRNAs have aroused general interest from exponentially increasing transcriptomic repertoires reporting their highly tissue-specific and developmentally dynamic expression, and more importantly, from growing experimental evidence supporting their functionality in facilitating organogenesis and individual fitness. In mammalian testes, while a great multitude of lncRNA species are identified, only a minority of them have been shown to be useful, and even fewer have been demonstrated as true requirements for male fertility using knockout models to date. This noticeable gap is attributed to the virtual existence of a large number of junk lncRNAs, the lack of an ideal germline culture system, difficulty in loss-of-function interrogation, and limited screening strategies. Facing these challenges, in this review, we discuss lncRNA functionality in organismal development and especially in mouse testis, with a focus on lncRNAs with functional screening.

LncRNA Functional Screening in Organismal Development.

Despite continuous expansion of the RNA binding protein (RBP) world, there is a lack of systematic understanding of RBPs in the mammalian testis, which harbors one of the most complex tissue transcriptomes. We adapted RNA interactome capture to mouse male germ cells, building an RBP atlas characterized by multiple layers of dynamics along spermatogenesis. Trapping of RNA-cross-linked peptides showed that the glutamic acid-arginine (ER) patch, a residue-coevolved polyampholytic element present in coiled coils, enhances RNA binding of its host RBPs. Deletion of this element in NONO (non-POU domain-containing octamer-binding protein) led to a defective mitosis-to-meiosis transition due to compromised NONO-RNA interactions. Whole-exome sequencing of over 1000 infertile men revealed a prominent role of RBPs in the human genetic architecture of male infertility and identified risk ER patch variants.

Article	Year
Microinjection of antisense oligonucleotides into living mouse testis enables lncRNA function study. Cell & bioscience\| PubMed ID: 34920761	2021
LncRNA Functional Screening in Organismal Development. Non-coding RNA\| PubMed ID: 37489456	2023
The landscape of RNA binding proteins in mammalian spermatogenesis. Science (New York, N.Y.)\| PubMed ID: 39208083	2024

Lingxiu Tong

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