scholarly journals New horizons in genome engineering of Drosophila melanogaster

2014 ◽  
Vol 89 (1) ◽  
pp. 3-8 ◽  
Author(s):  
Shu Kondo
Keyword(s):  
Drosophila Melanogaster ◽  
Genome Engineering ◽  
New Horizons

Genome editing in Drosophila melanogaster: from basic genome engineering to the multipurpose CRISPR-Cas9 system

2017 ◽  
Vol 60 (5) ◽  
pp. 476-489 ◽  
Author(s):  
Xingjie Ren ◽  
Kristof Holsteens ◽  
Haiyi Li ◽  
Jin Sun ◽  
Yifan Zhang ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Genome Editing ◽  
Genome Engineering ◽  
Basic Genome

scholarly journals Correction: Efficient multiplexed genome engineering with a polycistronic tRNA and CRISPR guide-RNA reveals an important role of detonator in reproduction of Drosophila melanogaster

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0250188
Author(s):  
Cristin Chon ◽  
Grace Chon ◽  
Yurika Matsui ◽  
Huiqing Zeng ◽  
Zhi-Chun Lai ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Genome Engineering ◽  
Guide Rna

scholarly journals Tissue-Specific DNA Replication Defects in Drosophila melanogaster Caused by a Meier-Gorlin Syndrome Mutation in Orc4

Genetics ◽  
2019 ◽  
Vol 214 (2) ◽  
pp. 355-367 ◽  
Author(s):  
Stephen L. McDaniel ◽  
Allison J. Hollatz ◽  
Anna M. Branstad ◽  
Marissa M. Gaskill ◽  
Catherine A. Fox ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Dna Replication ◽  
Origin Recognition Complex ◽  
Essential Role ◽  
Genome Engineering ◽  
Gorlin Syndrome ◽  
Developmental Defects ◽  
Tissue Specific ◽  
Link Type ◽  
Genes Encoding

Meier-Gorlin syndrome is a rare recessive disorder characterized by a number of distinct tissue-specific developmental defects. Genes encoding members of the origin recognition complex (ORC) and additional proteins essential for DNA replication (CDC6, CDT1, GMNN, CDC45, MCM5, and DONSON) are mutated in individuals diagnosed with MGS. The essential role of ORC is to license origins during the G1 phase of the cell cycle, but ORC has also been implicated in several nonreplicative functions. Because of its essential role in DNA replication, ORC is required for every cell division during development. Thus, it is unclear how the Meier-Gorlin syndrome mutations in genes encoding ORC lead to the tissue-specific defects associated with the disease. To begin to address these issues, we used Cas9-mediated genome engineering to generate a Drosophila melanogaster model of individuals carrying a specific Meier-Gorlin syndrome mutation in ORC4 along with control strains. Together these strains provide the first metazoan model for an MGS mutation in which the mutation was engineered at the endogenous locus along with precisely defined control strains. Flies homozygous for the engineered MGS allele reach adulthood, but with several tissue-specific defects. Genetic analysis revealed that this Orc4 allele was a hypomorph. Mutant females were sterile, and phenotypic analyses suggested that defects in DNA replication was an underlying cause. By leveraging the well-studied Drosophila system, we provide evidence that a disease-causing mutation in Orc4 disrupts DNA replication, and we propose that in individuals with MGS defects arise preferentially in tissues with a high-replication demand.

scholarly journals Programmable RNA Targeting using CasRx in Flies

2020 ◽  
Author(s):  
Anna Buchman ◽  
Dan J. Brogan ◽  
Ruichen Sun ◽  
Ting Yang ◽  
Patrick Hsu ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Genome Engineering ◽  
Developmental Stages ◽  
Comprehensive Evaluation ◽  
Model Organism ◽  
Endogenous Expression ◽  
Current State ◽  
Rna Targeting ◽  
Transcript Cleavage ◽  
Early Developmental

AbstractCRISPR-Cas genome editing technologies have revolutionized the fields of functional genetics and genome engineering, but with the recent discovery and optimization of RNA-targeting Cas ribonucleases, we may soon see a similar revolution in the study of RNA function and transcriptome engineering. However, to date, successful proof-of-principle for Cas ribonuclease RNA targeting in eukaryotic systems has been limited. Only recently has successful modification of RNA expression by a Cas ribonuclease been demonstrated in animal embryos. This previous work, however, did not evaluate endogenous expression of Cas ribonucleases and only focused on Cas ribonuclease function in early developmental stages. A more comprehensive evaluation of this technology is needed to assess its potential impact in the field. Here we report on our efforts to develop a programmable platform for RNA-targeting using a Cas ribonuclease, CasRx, in the model organism Drosophila melanogaster. By genetically encoding CasRx in flies, we demonstrate moderate transcript targeting of known phenotypic genes in addition to unexpected toxicity and lethality. We also report on the off-target effects following on-target transcript cleavage by CasRx. Taken together, our results present the current state and limitations of a genetically encoded programmable RNA-targeting Cas system in Drosophila melanogaster, paving the way for future optimization of the system.

scholarly journals Efficient multiplexed genome engineering with a polycistronic tRNA and CRISPR guide-RNA reveals an important role of detonator in reproduction of Drosophila melanogaster

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0245454
Author(s):  
Cristin Chon ◽  
Grace Chon ◽  
Yurika Matsui ◽  
Huiqing Zeng ◽  
Zhi-Chun Lai ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Genome Engineering ◽  
Transmembrane Protein ◽  
Association Studies ◽  
Fruit Fly ◽  
Loss Of Function ◽  
Guide Rna ◽  
Genome Association

Genome association studies in human and genetic studies in mouse implicated members of the transmembrane protein 132 (TMEM132) family in multiple conditions including panic disorder, hearing loss, limb and kidney malformation. However, the presence of five TMEM132 paralogs in mammalian genomes makes it extremely challenging to reveal the full requirement for these proteins in vivo. In contrast, there is only one TMEM132 homolog, detonator (dtn), in the genome of fruit fly Drosophila melanogaster, enabling straightforward research into its in vivo function. In the current study, we generate multiple loss-of-function dtn mutant fly strains through a polycistronic tRNA-gRNA approach, and show that most embryos lacking both maternal and paternal dtn fail to hatch into larvae, indicating an essential role of dtn in Drosophila reproduction.

scholarly journals A Meier-Gorlin Syndrome Mutation in Orc4 Causes Tissue-Specific DNA Replication Defects in Drosophila melanogaster

2019 ◽  
Author(s):  
Stephen L. McDaniel ◽  
Anna M. Branstad ◽  
Allison J. Hollatz ◽  
Catherine A. Fox ◽  
Melissa M. Harrison
Keyword(s):  
Drosophila Melanogaster ◽  
Dna Replication ◽  
Origin Recognition Complex ◽  
Genome Engineering ◽  
Primary Role ◽  
Gorlin Syndrome ◽  
Developmental Defects ◽  
Tissue Specific ◽  
Heterochromatin Formation ◽  
Genes Encoding

AbstractMeier-Gorlin syndrome is a rare recessive disorder characterized by a number of distinct developmental defects, including primordial dwarfism, small ears, and small or missing patella. Genes encoding members of the origin recognition complex (ORC) and additional proteins essential for DNA replication (CDC6, CDT1, GMNN, CDC45, and MCM5) are mutated in individuals diagnosed with MGS. The primary role of ORC is to license origins during the G1 phase of the cell cycle, but it also plays roles in cilia development, heterochromatin formation, and other cellular processes. Because of its essential role in DNA replication, ORC is required for every cell division during development. Thus, it is unclear how the Meier-Gorlin syndrome mutations in ORC lead to the tissue-specific defects associated with the disease. To address this question, we have used Cas9-mediated genome engineering to generate a Drosophila melanogaster model of individuals carrying a mutation in ORC4. Like the people with Meier-Gorlin syndrome, these flies reach adulthood, but have several tissue-specific defects. Genetic analysis revealed that this allele is a hypomorph and that mutant females are sterile. We demonstrated that this sterility is caused by a failure in DNA replication. By leveraging the well-studied Drosophila system, we showed that a disease-causing mutation in orc4 disrupts DNA replication, and we propose that in individuals with MGS defects arise preferentially in tissues with a high-replication demand.

Cas9-Mediated Genome Engineering in Drosophila melanogaster

2016 ◽  
Vol 2016 (9) ◽  
pp. pdb.top086843 ◽  
Author(s):  
Benjamin E. Housden ◽  
Norbert Perrimon
Keyword(s):  
Drosophila Melanogaster ◽  
Genome Engineering
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