University of Maryland Institute for Bioscience and Biotechnology Research

2 ARTICLES PUBLISHED IN JoVE

Genetics

Sand Fly (Phlebotomus papatasi) Embryo Microinjection for CRISPR/Cas9 Mutagenesis

Isabelle Louradour¹, Kashinath Ghosh¹, Ehud Inbar¹, David L. Sacks¹, Channa Aluvihare², Robert A. Harrell II²

¹Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, ²University of Maryland Insect Transformation Facility, The Institute for Bioscience and Biotechnology Research

This protocol details the steps of CRISPR/Cas9 targeted mutagenesis in sand flies: embryo collection, injection, insect rearing, and identification as well as selection of mutations of interest.

Biology

Embryo Injection Technique for Gene Editing in the Black-Legged Tick, Ixodes scapularis

Arvind Sharma¹, Michael Pham¹, Robert A. Harrell II², Andrew B. Nuss³, Monika Gulia-Nuss¹

¹Department of Biochemistry and Molecular Biology, University of Nevada, Reno, ²Insect Transformation Facility, University of Maryland Institute for Bioscience and Biotechnology Research, ³Department of Agriculture, Veterinary, and Rangeland Sciences, University of Nevada, Reno

The present protocol describes a method for injecting tick embryos. Embryo injection is the preferred technique for genetic manipulation to generate transgenic lines.