Your browser doesn't support javascript.
loading
A selectable, plasmid-based system to generate CRISPR/Cas9 gene edited and knock-in mosquito cell lines.
Rozen-Gagnon, Kathryn; Yi, Soon; Jacobson, Eliana; Novack, Sasha; Rice, Charles M.
Afiliación
  • Rozen-Gagnon K; Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY, 10065, USA. krozen@rockefeller.edu.
  • Yi S; Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY, 10065, USA.
  • Jacobson E; Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY, 10065, USA.
  • Novack S; Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY, 10065, USA.
  • Rice CM; Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY, 10065, USA.
Sci Rep ; 11(1): 736, 2021 01 12.
Article en En | MEDLINE | ID: mdl-33436886
ABSTRACT
Aedes (Ae.) aegypti and Ae. albopictus mosquitoes transmit arthropod-borne diseases around the globe, causing ~ 700,000 deaths each year. Genetic mutants are valuable tools to interrogate both fundamental vector biology and mosquito host factors important for viral infection. However, very few genetic mutants have been described in mosquitoes in comparison to model organisms. The relative ease of applying CRISPR/Cas9-based gene editing has transformed genome engineering and has rapidly increased the number of available gene mutants in mosquitoes. Yet, in vivo studies may not be practical for screening large sets of mutants or possible for laboratories that lack insectaries. Thus, it would be useful to adapt CRISPR/Cas9 systems to common mosquito cell lines. In this study, we generated and characterized a mosquito optimized, plasmid-based CRISPR/Cas9 system for use in U4.4 (Ae. albopictus) and Aag2 (Ae. aegypti) cell lines. We demonstrated highly efficient editing of the AGO1 locus and isolated U4.4 and Aag2 cell lines with reduced AGO1 expression. Further, we used homology-directed repair to establish knock-in Aag2 cell lines with a 3xFLAG-tag at the N-terminus of endogenous AGO1. These experimentally verified plasmids are versatile, cost-effective, and efficiently edit immune competent mosquito cell lines that are widely used in arbovirus studies.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Contexto en salud: 3_ND / 4_TD Problema de salud: 3_dengue / 4_dengue Asunto principal: Plásmidos / Aedes / Factores Eucarióticos de Iniciación / Sistemas CRISPR-Cas / Edición Génica / Mosquitos Vectores Tipo de estudio: Prognostic_studies Límite: Animals Idioma: En Revista: Sci Rep Año: 2021 Tipo del documento: Article País de afiliación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Contexto en salud: 3_ND / 4_TD Problema de salud: 3_dengue / 4_dengue Asunto principal: Plásmidos / Aedes / Factores Eucarióticos de Iniciación / Sistemas CRISPR-Cas / Edición Génica / Mosquitos Vectores Tipo de estudio: Prognostic_studies Límite: Animals Idioma: En Revista: Sci Rep Año: 2021 Tipo del documento: Article País de afiliación: Estados Unidos
...