scholarly journals VIGS-Mediated Forward Genetics Screening for Identification of Genes Involved in Nonhost Resistance

10.3791/51033 ◽  
2013 ◽  
Author(s):  
Muthappa Senthil-Kumar ◽  
Hee-Kyung Lee ◽  
Kirankumar S. Mysore
Keyword(s):  
Forward Genetics ◽  
Nonhost Resistance ◽  
Genetics Screening

Forward Genetics Screening of Medicago truncatula Tnt1 Insertion Lines

2013 ◽  
pp. 93-100 ◽  
Author(s):  
Juan Carlos Serrani Yarce ◽  
Hee-Kyung Lee ◽  
Million Tadege ◽  
Pascal Ratet ◽  
Kirankumar S. Mysore
Keyword(s):  
Medicago Truncatula ◽  
Forward Genetics ◽  
Genetics Screening

scholarly journals FORWARD AND UNBIASED RNA-INTERFERENCE SCREEN

2021 ◽  
Author(s):  
Wadim J Kapulkin
Keyword(s):  
Rna Interference ◽  
Caenorhabditis Elegans ◽  
Reverse Genetics ◽  
Screening Strategy ◽  
Rnai Screen ◽  
Forward Genetics ◽  
Genome Wide ◽  
Complementary Resource ◽  
Genetics Screening ◽  
Caenorhabditis Elegans Genome

RNA-interference (Fire et al. 1998) is a popular ‘reverse-genetics’ screening strategy applied in Caenorhabditis elegans. Genome-wide RNAi screens are presently carried using RNAi feeding libraries. Here, we report on a complementary resource facilitating an approach to RNAi screen relying on an unbiased ‘forward-genetics’ strategy. We conclude the forward RNA interference screening is useful and feasible, with the strong expectation the presented screening mode will complement and extend on the existing, currently available, genome-wide RNAi resources.

scholarly journals Large-scale forward genetics screening identifies Trpa1 as a chemosensor for predator odor-evoked innate fear behaviors

2018 ◽  
Vol 9 (1) ◽  
Author(s):  
Yibing Wang ◽  
Liqin Cao ◽  
Chia-Ying Lee ◽  
Tomohiko Matsuo ◽  
Kejia Wu ◽  
...  
Keyword(s):  
Large Scale ◽  
Forward Genetics ◽  
Predator Odor ◽  
Genetics Screening ◽  
Innate Fear

scholarly journals A novel gene-diet interaction promotes organismal lifespan and host protection during infection via the mitochondrial UPR

PLoS Genetics ◽  
2020 ◽  
Vol 16 (12) ◽  
pp. e1009234
Author(s):  
Mustafi Raisa Amin ◽  
Siraje Arif Mahmud ◽  
Jonathan L. Dowgielewicz ◽  
Madhab Sapkota ◽  
Mark W. Pellegrino
Keyword(s):  
Vitamin B12 ◽  
Ribosomal Gene ◽  
Forward Genetics ◽  
Dependent Manner ◽  
Unfolded Protein ◽  
Host Protection ◽  
Novel Gene ◽  
Extended Lifespan ◽  
Protein Response ◽  
Genetics Screening

Cells use a variety of mechanisms to maintain optimal mitochondrial function including the mitochondrial unfolded protein response (UPRmt). The UPRmt mitigates mitochondrial dysfunction by differentially regulating mitoprotective gene expression through the transcription factor ATFS-1. Since UPRmt activation is commensurate with organismal benefits such as extended lifespan and host protection during infection, we sought to identify pathways that promote its stimulation. Using unbiased forward genetics screening, we isolated novel mutant alleles that could activate the UPRmt. Interestingly, we identified one reduction of function mutant allele (osa3) in the mitochondrial ribosomal gene mrpl-2 that activated the UPRmt in a diet-dependent manner. We find that mrpl-2(osa3) mutants lived longer and survived better during pathogen infection depending on the diet they were fed. A diet containing low levels of vitamin B12 could activate the UPRmt in mrpl-2(osa3) animals. Also, we find that the vitamin B12-dependent enzyme methionine synthase intersects with mrpl-2(osa3) to activate the UPRmt and confer animal lifespan extension at the level of ATFS-1. Thus, we present a novel gene-diet pairing that promotes animal longevity that is mediated by the UPRmt.

scholarly journals A PCR-based forward genetics screening, using expression domain-specific markers, identifies mutants in endosperm transfer cell development

2014 ◽  
Vol 5 ◽  
Author(s):  
Luis M. Muñiz ◽  
Elisa Gómez ◽  
Virginie Guyon ◽  
Maribel López ◽  
Bouchaib Khbaya ◽  
...  
Keyword(s):  
Cell Development ◽  
Forward Genetics ◽  
Transfer Cell ◽  
Expression Domain ◽  
Domain Specific ◽  
Endosperm Transfer Cell ◽  
Genetics Screening

scholarly journals Massively parallel in vivo CRISPR screening identifies RNF20/40 as epigenetic regulators of cardiomyocyte maturation

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Nathan J. VanDusen ◽  
Julianna Y. Lee ◽  
Weiliang Gu ◽  
Catalina E. Butler ◽  
Isha Sethi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Genetic Screen ◽  
Forward Genetics ◽  
Epigenetic Mark ◽  
Biological Processes ◽  
Dynamic Changes ◽  
Forward Genetic Screen ◽  
High Resource ◽  
Organ Systems

AbstractThe forward genetic screen is a powerful, unbiased method to gain insights into biological processes, yet this approach has infrequently been used in vivo in mammals because of high resource demands. Here, we use in vivo somatic Cas9 mutagenesis to perform an in vivo forward genetic screen in mice to identify regulators of cardiomyocyte (CM) maturation, the coordinated changes in phenotype and gene expression that occur in neonatal CMs. We discover and validate a number of transcriptional regulators of this process. Among these are RNF20 and RNF40, which form a complex that monoubiquitinates H2B on lysine 120. Mechanistic studies indicate that this epigenetic mark controls dynamic changes in gene expression required for CM maturation. These insights into CM maturation will inform efforts in cardiac regenerative medicine. More broadly, our approach will enable unbiased forward genetics across mammalian organ systems.

scholarly journals Fast genetic mapping using insertion-deletion polymorphisms in Caenorhabditis elegans

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ho-Yon Hwang ◽  
Jiou Wang
Keyword(s):  
Caenorhabditis Elegans ◽  
Genetic Mapping ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Genetic Material ◽  
Mapping Method ◽  
Forward Genetics ◽  
Whole Genome ◽  
Nucleotide Polymorphisms ◽  
Large Populations

AbstractGenetic mapping is used in forward genetics to narrow the list of candidate mutations and genes corresponding to the mutant phenotype of interest. Even with modern advances in biology such as efficient identification of candidate mutations by whole-genome sequencing, mapping remains critical in pinpointing the responsible mutation. Here we describe a simple, fast, and affordable mapping toolkit that is particularly suitable for mapping in Caenorhabditis elegans. This mapping method uses insertion-deletion polymorphisms or indels that could be easily detected instead of single nucleotide polymorphisms in commonly used Hawaiian CB4856 mapping strain. The materials and methods were optimized so that mapping could be performed using tiny amount of genetic material without growing many large populations of mutants for DNA purification. We performed mapping of previously known and unknown mutations to show strengths and weaknesses of this method and to present examples of completed mapping. For situations where Hawaiian CB4856 is unsuitable, we provide an annotated list of indels as a basis for fast and easy mapping using other wild isolates. Finally, we provide rationale for using this mapping method over other alternatives as a part of a comprehensive strategy also involving whole-genome sequencing and other methods.

scholarly journals Toward genetic modification of plant-parasitic nematodes: delivery of macromolecules to adults and expression of exogenous mRNA in second stage juveniles

2021 ◽  
Vol 11 (2) ◽  
Author(s):  
Olaf Kranse ◽  
Helen Beasley ◽  
Sally Adams ◽  
Andre Pires-daSilva ◽  
Christopher Bell ◽  
...  
Keyword(s):  
Food Security ◽  
Genetic Modification ◽  
The Body ◽  
Forward Genetics ◽  
Heterodera Schachtii ◽  
Parasitic Nematodes ◽  
Plant Parasitic Nematodes ◽  
Root Knot Nematode ◽  
Cyst Nematodes ◽  
Plant Parasitic

Abstract Plant-parasitic nematodes are a continuing threat to food security, causing an estimated 100 billion USD in crop losses each year. The most problematic are the obligate sedentary endoparasites (primarily root knot nematodes and cyst nematodes). Progress in understanding their biology is held back by a lack of tools for functional genetics: forward genetics is largely restricted to studies of natural variation in populations and reverse genetics is entirely reliant on RNA interference. There is an expectation that the development of functional genetic tools would accelerate the progress of research on plant-parasitic nematodes, and hence the development of novel control solutions. Here, we develop some of the foundational biology required to deliver a functional genetic tool kit in plant-parasitic nematodes. We characterize the gonads of male Heterodera schachtii and Meloidogyne hapla in the context of spermatogenesis. We test and optimize various methods for the delivery, expression, and/or detection of exogenous nucleic acids in plant-parasitic nematodes. We demonstrate that delivery of macromolecules to cyst and root knot nematode male germlines is difficult, but possible. Similarly, we demonstrate the delivery of oligonucleotides to root knot nematode gametes. Finally, we develop a transient expression system in plant-parasitic nematodes by demonstrating the delivery and expression of exogenous mRNA encoding various reporter genes throughout the body of H. schachtii juveniles using lipofectamine-based transfection. We anticipate these developments to be independently useful, will expedite the development of genetic modification tools for plant-parasitic nematodes, and ultimately catalyze research on a group of nematodes that threaten global food security.

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