scholarly journals Development of zygotic and germline gene drives in mice

2020 ◽  
Author(s):  
Chandran Pfitzner ◽  
James Hughes ◽  
Melissa White ◽  
Michaela Scherer ◽  
Sandra Piltz ◽  
...  
Keyword(s):  
Pest Control ◽  
Mus Musculus ◽  
Synthetic Gene ◽  
Germline Gene ◽  
Intended Target ◽  
Vertebrate Pest ◽  
Critical Requirement ◽  
Drive Systems ◽  
Gene Drives

AbstractCRISPR-based synthetic gene drives have the potential to deliver a more effective and humane method of invasive vertebrate pest control than current strategies. Relatively efficient CRISPR gene drives have been developed in insects and yeast but not in mammals. Here we investigated the efficiency of CRISPR/Cas9-based gene drives in Mus musculus by constructing “split drive” systems with Cas9 under the control of zygotic (CAG) or germline (Vasa) promoters. While both systems generated double stranded breaks at their intended target site in vivo, no homing was detectable. Our data indicate that robust and specific Cas9 expression during meiosis is a critical requirement for the generation of efficient CRISPR-based synthetic gene drives in rodents.

scholarly journals Overcoming evolved resistance to population-suppressing homing-based gene drives

2016 ◽  
Author(s):  
John M. Marshall ◽  
Anna Buchman ◽  
Héctor M. Sánchez C. ◽  
Omar S. Akbari
Keyword(s):  
Mosquito Species ◽  
Gene Drive ◽  
Resistant Allele ◽  
Guide Rnas ◽  
Continental Scale ◽  
Fitness Advantage ◽  
Health Related ◽  
Drive Systems ◽  
Gene Drives

AbstractThe use of homing-based gene drive systems to modify or suppress wild populations of a given species has been proposed as a solution to a number of significant ecological and public health related problems, including the control of mosquito-borne diseases. The recent development of a CRISPR-Cas9-based homing system for the suppression ofAnopheles gambiae, the main African malaria vector, is encouraging for this approach; however, with current designs, the slow emergence of homing-resistant alleles is expected to result in suppressed populations rapidly rebounding, as homing-resistant alleles have a significant fitness advantage over functional, population-suppressing homing alleles. To explore this concern, we develop a mathematical model to estimate tolerable rates of homing-resistant allele generation to suppress a wild population of a given size. Our results suggest that, to achieve meaningful population suppression, tolerable rates of resistance allele generation are orders of magnitude smaller than those observed for current designs for CRISPR-Cas9-based homing systems. To remedy this, we propose a homing system architecture in which guide RNAs (gRNAs) are multiplexed, increasing the effective homing rate and decreasing the effective resistant allele generation rate. Modeling results suggest that the size of the population that can be suppressed increases exponentially with the number of multiplexed gRNAs and that, with six multiplexed gRNAs, a mosquito species could potentially be suppressed on a continental scale. We also demonstrate successful multiplexingin vivoinDrosophila melanogasterusing a ribozyme-gRNA-ribozyme (RGR) approach – a strategy that could readily be adapted to engineer stable, homing-based suppression drives in relevant organisms.

EFECTOS CITOGENÉTICOS DE LA FÓRMULA COMERCIAL DE UNA TABLETA ANTIGRIPAL EN LOS ERITROCITOS DE SANGRE PERIFÉRICA DE RATÓN ÁRABE (MUS MUSCULUS LINNAEUS, 1758)

2013 ◽  
Vol 6 ◽  
Author(s):  
Saúl Flores Maya ◽  
Héctor Barrera Escorcia ◽  
Alexis Frausto Cornejo ◽  
Daniela Elizabeth Chávez Vázquez ◽  
Ana Cristina Hernández Cruz ◽  
...  
Keyword(s):  
Mus Musculus

La mezcla de analgésico, antipirético y antihistamínico de una pastilla antigripal de marca conocida fue evaluada en su capacidad de provocar daño cromosómico y citotoxicidad en sangre periférica de ratón árabe. El daño cromosómico fue evaluado utilizando la prueba de micronúcleos in vivo. El antigripal fue administrado a ratones de la línea árabe por vía oral en una dosis de 10.6 mg/Kg de peso en el curso de ocho h por tres días en un tratamiento agudo. Los cálculos del índice de toxicidad no fueron significativos estadísticamente entre los datos del grupo control negativo y los tratamientos con el antigripal. En cambio, este antigripal mostró efectos genotóxicos significativos a las 24, 48 y 72 h después de su aplicación. Este efecto puede ser causa de los componentes químicos del antigripal como son el paracetamol y la cafeína. En conclusión, la mezcla de analgésico, antipirético y antihistamínico de una pastilla antigripal no provoca daño celular pero muestra daño clastogénico durante el tratamiento agudo del antigripal en ratones de la línea árabe. El ratón Árabe mostró sensibilidad a los efectos de los agentes genotóxicos y, por tanto, este organismo debería ser incluido para estudios de genotoxicidad.

scholarly journals Pengaruh Pemberian Taurin terhadap Gambaran Histopatologi Paru Mencit (Mus musculus) yang Diinduksi Karsinogen Benzo(α)Piren secara In Vivo

2017 ◽  
Vol 17 (1) ◽  
pp. 22
Author(s):  
Arini Pradita Roselyn ◽  
Endang Linirin Widiastuti ◽  
G. Nugroho Susanto ◽  
Sutyarso '
Keyword(s):  
Lung Tissue ◽  
Mus Musculus ◽  
Tissue Damage ◽  
Corn Oil ◽  
Group Vi ◽  
Normal Tissues ◽  
Randomized Design ◽  
Group I ◽  
Lung Tissue Damage

Lung cancer is a disease that causes high mortality. Drugs used to prevent and cure cancer mostly causes intoxicity to the normal tissues due to its less effectiveness. Therefore, it is necessary to find out any agent or substance which works much more effective and safe for cancer treatment. The aim of the study was to elucidate the role of taurine on the lung tissue of mice (Mus musculus) induced by carcinogenic benzo(α)pyrene. The experiment was conducted in a completely randomized design with 5 replications. Six treatment groups were perfomed. Group I was given 0.2 mL of corn oil and given aquadest until the end of the study period, group II was induced by benzo(α)pyrene without administration of taurine, group III before induced with benzo(α)pyrene, was given taurine dosage 7.8 mg/BW/day for two weeks, group IV after induced benzo(α)pyrene, was given taurine with dosage 3.9 mg/BW/day, group V after induced benzo(α)pyrene, was given taurine with dosage7. 8 mg/BW/day, group VI after induced with benzo(α)pyrene, was given taurine with dosage 15.6 mg/BW/day. The results of the Kruskal-Wallis analysis and one way ANOVA with LSD (p>0,05) showed that taurine reduced lung tissue damage 72.73% due to the administration of benzo(α)pyrene of 0.3 mg/BW/day. In addition, the effective dose of taurine reduce lung tissue damage was 15.6 mg/BW/day.

scholarly journals EFECTO FOTOPROTECTOR DE Macrocystis pyrifera FRENTE A BLOQUEADORES SOLARES EN LA PREVENCIÓN DE LESIONES DE PIEL

2015 ◽  
Vol 18 (2) ◽  
pp. 95-98
Author(s):  
Américo J. Castro ◽  
Mario Carhuapoma ◽  
Norma J. Ramos ◽  
José R. Juárez ◽  
Luis M. Felix ◽  
...  
Keyword(s):  
Mus Musculus ◽  
Macrocystis Pyrifera ◽  
San Juan ◽  
Anterior Dorsal ◽  
Los Grupos

El estudio tuvo como objetivo comparar el efecto fotoprotector in vivo de una crema elaborada con el extracto hidroalcohólico de la macroalga parda Macrocystis pyrifera (Linnapus) Agardh frente a bloqueadores solares comerciales en la prevención de lesiones de piel. La muestra biológica fue colectada en el zócalo continental de la playa Yanyarina de San Juan de Marcona de la región Ica. El extracto hidroalcohólico (etanol-agua 1:1) se obtuvo por proceso de maceración a partir de 300 g de alga. Con el extracto se formuló y diseño el bloqueador dermocosmético empleando concentraciones 5, 10 y 15%, respectivamente, con una mezcla de alcoholes alifáticos sólidos, conformados principalmente por alcohol cetílico (C16H3O) y alcohol estearílico (C18H39O) 50/70%. Se emplearon 35 ratones albinos adultos machos de la especie Mus musculus cepa Bald C53 de peso promedio 30g. El potencial fotoprotector in vivo se evaluó a través de la aplicación tópica en la piel de los ratones, en el tercio anterior dorsal del lomo. Los animales fueron distribuidos en siete grupos de cinco. Al grupo 1 (blanco) no se le aplicó crema ni irradiación UV-B 320 ŋm; al grupo 2 (control) no se le aplicó crema y se le irradió con luz UV-B; los grupos 3 y 4 fueron tratados con bloqueadores solares comerciales y fueron irradiados con luz UV-B; a los grupos 5, 6 y 7 (intervención) se les aplicó el producto elaborado y se le irradió con luz UV-B. El efecto fotoprotector se evidenció en el análisis macroscópico y microscópico de la piel. En el análisis microscópico, el grupo 1 presento piel con estructura conservada; los grupos 2, 3 y 4 presentaron estructura alterada; mientras que el grupo 5 presentó alteración leve, el grupo 6, estructura aparentemente normal y el grupo 7 estructura normal. Se concluye que el extracto hidroalcohólico ejerce efecto fotoprotector in vivo ante la irradiación UV-B en las concentraciones trabajadas, lográndose que, a medida que se va aumentando la concentración del extracto, la estructura de la piel se ve menos alterada presentando la dermis y epidermis, la capa cornea laminar y basal aparentemente normales.

Containment strategies for synthetic gene drive organisms and impacts on gene flow.

2021 ◽  
pp. 137-152
Author(s):  
Lei Pei ◽  
Markus Schmidt
Keyword(s):  
Gene Flow ◽  
Fungal Infections ◽  
Basic Research ◽  
Fruit Fly ◽  
Mitigation Strategies ◽  
Synthetic Gene ◽  
Model Organisms ◽  
Gene Drive ◽  
Comprehensive Overview ◽  
Gene Drives

Abstract Gene drives, particularly synthetic gene drives, may help to address some important challenges, by efficiently altering specific sections of DNA in entire populations of wild organisms. Here we review the current development of the synthetic gene drives, especially those RNA-guided synthetic gene drives based on the CRISPR nuclease Cas. Particular focuses are on their possible applications in agriculture (e.g. disease resistance, weed control management), ecosystem conservation (e.g. evasion species control), health (e.g. to combat insect-borne and fungal infections), and for basic research in model organisms (e.g. Saccharomyces, fruit fly, and zebra fish). The physical, chemical, biological, and ecological containment strategies that might help to confine these gene drive-modified organisms are then explored. The gene flow issues, those from gene drive-derived organisms to the environment, are discussed, while possible mitigation strategies for gene drive research are explored. Last but not least, the regulatory context and opinions from key stakeholders (regulators, scientists, and concerned organizations) are reviewed, aiming to provide a more comprehensive overview of the field.

scholarly journals Environmental RNA interference in two-spotted spider mite, Tetranychus urticae, reveals dsRNA processing requirements for efficient RNAi response

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Nicolas Bensoussan ◽  
Sameer Dixit ◽  
Midori Tabara ◽  
David Letwin ◽  
Maja Milojevic ◽  
...  
Keyword(s):  
Rna Interference ◽  
Tetranychus Urticae ◽  
Pest Control ◽  
Spider Mite ◽  
26S Proteasome ◽  
Rnai Pathway ◽  
Rnai Machinery ◽  
Two Spotted Spider Mite ◽  
Species Specific

Abstract Comprehensive understanding of pleiotropic roles of RNAi machinery highlighted the conserved chromosomal functions of RNA interference. The consequences of the evolutionary variation in the core RNAi pathway genes are mostly unknown, but may lead to the species-specific functions associated with gene silencing. The two-spotted spider mite, Tetranychus urticae, is a major polyphagous chelicerate pest capable of feeding on over 1100 plant species and developing resistance to pesticides used for its control. A well annotated genome, susceptibility to RNAi and economic importance, make T. urticae an excellent candidate for development of an RNAi protocol that enables high-throughput genetic screens and RNAi-based pest control. Here, we show that the length of the exogenous dsRNA critically determines its processivity and ability to induce RNAi in vivo. A combination of the long dsRNAs and the use of dye to trace the ingestion of dsRNA enabled the identification of genes involved in membrane transport and 26S proteasome degradation as sensitive RNAi targets. Our data demonstrate that environmental RNAi can be an efficient reverse genetics and pest control tool in T. urticae. In addition, the species-specific properties together with the variation in the components of the RNAi machinery make T. urticae a potent experimental system to study the evolution of RNAi pathways.

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