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      mRNA localization in the Drosophila germline

      review-article
      Author(s): *
      Publication date (Electronic):
      Journal: RNA Biology
      Publisher: Taylor & Francis
      Keywords: mRNA localization, Drosophila germline, ribonucleoprotein (RNP) complex, active transport

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          Abstract

          Localization and the associated translational control of mRNA is a well established mechanism for segregating cellular protein expression. Drosophila has been instrumental in deciphering the prevailing mechanisms of mRNA localization and regulation. This review will discuss the diverse roles of mRNA localization in the Drosophila germline, the cis-elements and cellular components regulating localization and the superimposition of translational regulatory mechanisms. Despite a history of discovery, there are still many fundamental questions regarding mRNA localization that remain unanswered. Take home messages, outstanding questions and future approaches that will likely lead to resolving these unknowns in the future are summarized at the end.

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          Optimized CRISPR/Cas tools for efficient germline and somatic genome engineering in Drosophila.

          Fillip Port, Hui-Min Chen, Tzumin Lee … (2014)
          The type II clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system has emerged recently as a powerful method to manipulate the genomes of various organisms. Here, we report a toolbox for high-efficiency genome engineering of Drosophila melanogaster consisting of transgenic Cas9 lines and versatile guide RNA (gRNA) expression plasmids. Systematic evaluation reveals Cas9 lines with ubiquitous or germ-line-restricted patterns of activity. We also demonstrate differential activity of the same gRNA expressed from different U6 snRNA promoters, with the previously untested U6:3 promoter giving the most potent effect. An appropriate combination of Cas9 and gRNA allows targeting of essential and nonessential genes with transmission rates ranging from 25-100%. We also demonstrate that our optimized CRISPR/Cas tools can be used for offset nicking-based mutagenesis. Furthermore, in combination with oligonucleotide or long double-stranded donor templates, our reagents allow precise genome editing by homology-directed repair with rates that make selection markers unnecessary. Last, we demonstrate a novel application of CRISPR/Cas-mediated technology in revealing loss-of-function phenotypes in somatic cells following efficient biallelic targeting by Cas9 expressed in a ubiquitous or tissue-restricted manner. Our CRISPR/Cas tools will facilitate the rapid evaluation of mutant phenotypes of specific genes and the precise modification of the genome with single-nucleotide precision. Our results also pave the way for high-throughput genetic screening with CRISPR/Cas.
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            RNA and disease.

            Thomas Cooper, Lili Wan, Gideon Dreyfuss (2009)
            Cellular functions depend on numerous protein-coding and noncoding RNAs and the RNA-binding proteins associated with them, which form ribonucleoprotein complexes (RNPs). Mutations that disrupt either the RNA or protein components of RNPs or the factors required for their assembly can be deleterious. Alternative splicing provides cells with an exquisite capacity to fine-tune their transcriptome and proteome in response to cues. Splicing depends on a complex code, numerous RNA-binding proteins, and an enormously intricate network of interactions among them, increasing the opportunity for exposure to mutations and misregulation that cause disease. The discovery of disease-causing mutations in RNAs is yielding a wealth of new therapeutic targets, and the growing understanding of RNA biology and chemistry is providing new RNA-based tools for developing therapeutics.
            Article 0 comments Cited 368 times – based on 0 reviews     Review now
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              Global analysis of mRNA localization reveals a prominent role in organizing cellular architecture and function.

              Eric Lecuyer, Hideki Yoshida, Neela Parthasarathy … (2007)
              Although subcellular mRNA trafficking has been demonstrated as a mechanism to control protein distribution, it is generally believed that most protein localization occurs subsequent to translation. To address this point, we developed and employed a high-resolution fluorescent in situ hybridization procedure to comprehensively evaluate mRNA localization dynamics during early Drosophila embryogenesis. Surprisingly, of the 3370 genes analyzed, 71% of those expressed encode subcellularly localized mRNAs. Dozens of new and striking localization patterns were observed, implying an equivalent variety of localization mechanisms. Tight correlations between mRNA distribution and subsequent protein localization and function, indicate major roles for mRNA localization in nucleating localized cellular machineries. A searchable web resource documenting mRNA expression and localization dynamics has been established and will serve as an invaluable tool for dissecting localization mechanisms and for predicting gene functions and interactions.
              Article 0 comments Cited 326 times – based on 0 reviews     Review now
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                Author and article information

                Journal
                Journal ID (nlm-ta): RNA Biol
                Journal ID (iso-abbrev): RNA Biol
                Journal ID (pmc): KRNB
                Title: RNA Biology
                Publisher: Taylor & Francis
                ISSN (Print): 1547-6286
                ISSN (Electronic): 1555-8584
                Publication date Collection: August 2014
                Publication date (Electronic): 31 October 2014
                Volume: 11
                Issue: 8
                Pages: 1010-1018
                Affiliations
                Department of Zoology; University of Cambridge ; Cambridge, UK
                Author notes
                [* ]Correspondence to: Timothy T Weil; Email: tw419@ 123456cam.ac.uk
                Article
                Publisher ID: 972211
                DOI: 10.4161/rna.36097
                PMC ID: 4615827
                PubMed ID: 25482896
                SO-VID: 166a2f5e-99fb-4f50-a55d-b1e7209639d6
                Copyright © © 2014 The Author(s). Published with license by Taylor & Francis Group, LLC© Timothy T Weil
                License:

                This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

                History
                Date received : 8 August 2014
                Date revision received : 14 August 2014
                Date accepted : 15 August 2014
                Page count
                Figures: 1, Tables: 0, References: 117, Pages: 9
                Categories
                Subject: Review

                ScienceOpen disciplines: Molecular biology
                Keywords: mrna localization,drosophila germline,ribonucleoprotein (rnp) complex,active transport
                Data availability:
                ScienceOpen disciplines: Molecular biology
                Keywords: mrna localization, drosophila germline, ribonucleoprotein (rnp) complex, active transport

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