<i>curled</i>Encodes the Drosophila Homolog of the Vertebrate Circadian Deadenylase Nocturnin

Grönke, Sebastian; Bickmeyer, Iris; Wunderlich, Roman; Jäckle, Herbert; Kühnlein, Ronald P.

doi:10.1534/genetics.109.105601

Cited by 30 publications

(39 citation statements)

References 76 publications

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“…We used our dual gRNA plasmid to make a transgenic line simultaneously expressing gRNAs targeting e or curled (cu), which causes a curled wing phenotype when mutated (23), and crossing that line to act-cas9 flies. All 120 of the analyzed act-cas9 gRNA-e gRNA-cu flies had large patches of dark cuticle, and 52% had one or two curled wings (Fig.…”

Section: Resultsmentioning

confidence: 99%

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

Port

Chen

Lee

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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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 germline-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 doublestranded 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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Section: Resultsmentioning

confidence: 99%

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

Port

Chen

Lee

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

1,012

1,198

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“…Disrupting cu or HR4 function via RNAi in the RG or the PG reduced the duration of larval development in both genotypes by 10 (Figure 4D) or 7 hr (data not shown), respectively. Drosophila CU is orthologous to vertebrate Nocturnin (NOC), a circadian rhythm effector protein (Grönke et al, 2009). Interestingly, NOC interacts with the nuclear receptor PPARγ to facilitate its translocation to the nucleus, raising the possibility that a similar interaction may occur between CU and HR4.…”

Section: Resultsmentioning

confidence: 99%

The Insect Prothoracic Gland as a Model for Steroid Hormone Biosynthesis and Regulation

Zeng

Yamanaka

et al. 2016

Cell Reports

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SUMMARY Steroid hormones are ancient signaling molecules found in vertebrates and insects alike. Both taxa show intriguing parallels with respect to how steroids function and how their synthesis is regulated. As such, insects are excellent models for studying universal aspects of steroid physiology. Here, we present a comprehensive genomic and genetic analysis of the principal steroid hormone-producing organs in two popular insect models, Drosophila and Bombyx. We identified 173 genes with previously unknown specific expression in steroid-producing cells, 15 of which had critical roles in development. The insect neuropeptide PTTH and its vertebrate counterpart ACTH both regulate steroid production, but molecular targets of these pathways remain poorly characterized. Identification of PTTH-dependent gene sets identified the nuclear receptor HR4 as a highly conserved target in both Drosophila and Bombyx. We consider this study to be a critical step toward understanding how steroid hormone production and release are regulated in all animal models.

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“…In addition to CCR4, encoded by the gene twin, the Drosophila genome also encodes three paralogs: Angel, Nocturnin (encoded by curled) (Grönke et al 2009), and 3635. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that twin, angel, and 3635 are expressed in S2 cells, but curled is not (data not shown).…”

Section: Association Of Nocturnin With the Ccr4-not Complexmentioning

confidence: 99%

Subunits of the Drosophila CCR4-NOT complex and their roles in mRNA deadenylation

Temme¹,

Zhang²,

Kremmer³

et al. 2010

RNA

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The CCR4-NOT complex is the main enzyme catalyzing the deadenylation of mRNA. We have investigated the composition of this complex in Drosophila melanogaster by immunoprecipitation with a monoclonal antibody directed against NOT1. The CCR4, CAF1 (=POP2), NOT1, NOT2, NOT3, and CAF40 subunits were associated in a stable complex, but NOT4 was not. Factors known to be involved in mRNA regulation were prominent among the other proteins coprecipitated with the CCR4-NOT complex, as analyzed by mass spectrometry. The complex was localized mostly in the cytoplasm but did not appear to be a major component of P bodies. Of the known CCR4 paralogs, Nocturnin was found associated with the subunits of the CCR4-NOT complex, whereas Angel and 3635 were not. RNAi experiments in Schneider cells showed that CAF1, NOT1, NOT2, and NOT3 are required for bulk poly(A) shortening and hsp70 mRNA deadenylation, but knock-down of CCR4, CAF40, and NOT4 did not affect these processes. Overexpression of catalytically dead CAF1 had a dominant-negative effect on mRNA decay. In contrast, overexpression of inactive CCR4 had no effect. We conclude that CAF1 is the major catalytically important subunit of the CCR4-NOT complex in Drosophila Schneider cells. Nocturnin may also be involved in mRNA deadenylation, whereas there is no evidence for a similar role of Angel and 3635.

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curledEncodes the Drosophila Homolog of the Vertebrate Circadian Deadenylase Nocturnin

Cited by 30 publications

References 76 publications

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

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

The Insect Prothoracic Gland as a Model for Steroid Hormone Biosynthesis and Regulation

Subunits of the Drosophila CCR4-NOT complex and their roles in mRNA deadenylation

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