Two isoforms of the essential C. elegans Argonaute CSR-1 differentially regulate sperm and oocyte fertility
Amanda Charlesworth,Uri Seroussi,Nicolas J. Lehrbach,Mathias S. Renaud,Adam E. Sundby,Ruxandra I. Molnar,Robert X Lao,Alexandra R. Willis,Jenna R. Woock,Matthew J. Aber,Annette J. Diao,Aaron W. Reinke,Gary Ruvkun,Julie M. Claycomb +13 more
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TLDR
In this article, the authors used CRISPR-Cas9 genome editing to introduce GFP::3xFLAG into the long and short (CSR-1a) isoforms of the Caenorhabditis elegans genome and found that CSR1a is expressed during spermatogenesis and in several somatic tissues, including the intestine.Abstract:
The Caenorhabditis elegans genome encodes nineteen functional Argonaute proteins that use 22G-RNAs, 26G-RNAs, miRNAs or piRNAs to regulate target transcripts. Only one Argonaute is essential under normal laboratory conditions: CSR-1. While CSR-1 has been studied widely, nearly all studies have overlooked the fact that the csr-1 locus encodes two isoforms. These isoforms differ by an additional 163 amino acids present in the N-terminus of CSR-1a. Using CRISPR-Cas9 genome editing to introduce GFP::3xFLAG into the long (CSR-1a) and short (CSR-1b) isoforms, we found that CSR-1a is expressed during spermatogenesis and in several somatic tissues, including the intestine. CSR-1b is expressed constitutively in the germline. small RNA sequencing of CSR-1 complexes shows that they interact with partly overlapping sets of 22G-RNAs. Phenotypic analyses reveal that the essential functions of csr-1 described in the literature coincide with CSR-1b, while CSR-1a plays tissue specific functions. During spermatogenesis, CSR-1a integrates into an sRNA regulatory network including ALG-3, ALG-4 and WAGO-10 that is necessary for fertility at 25°C. In the intestine, CSR-1a silences immunity and pathogen-responsive genes, and its loss results in improved survival from the pathogen Pseudomonas aeruginosa. Our findings functionally distinguish the CSR-1 isoforms and highlight the importance of studying each AGO isoform independently.read more
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Journal ArticleDOI
Systematic characterization of small RNAs associated with C. elegans Argonautes
TL;DR: In this paper , an expanded Argonaute family with 20 potentially functional members has been identified in Caenorhabditis elegans and their associated sRNAs were analyzed.
Journal ArticleDOI
Application of CRISPR/Cas Technology in Spermatogenesis Research and Male Infertility Treatment
TL;DR: The applications of gene editing technologies, especially CRISPR/Cas9, in deepening the understanding of the function of spermatogenesis-related genes and disease treatment are summarized.
Posted ContentDOI
The evolution of an RNA-based memory of self in the face of genomic conflict
Pinelopi Pliota,Hana Marvanova,Alevtina Koreshova,Yotam Kaufman,Polina O. Tikanova,Daniel Krogull,Andreas Hagmüller,Sonya A. Widen,Dominik Handler,Joseph F. Gokcezade,Peter Duchek,Julius Brennecke,Eyal Ben-David,Alejandro Burga +13 more
TL;DR: The results indicate that epigenetic licensing is likely a common mechanism that hinders the spread of selfish genes in wild populations while ensuring a lasting memory of self in the germline.
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