利用CRISPR/Cas9对微小隐孢子虫进行基因操作

Genetic Manipulation of Cryptosporidium parvum with CRISPR/Cas9.

作者信息

Sateriale Adam, Pawlowic Mattie, Vinayak Sumiti, Brooks Carrie, Striepen Boris

机构信息

Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.

Wellcome Trust Centre for Anti-Infectives Research, University of Dundee, Dundee, UK.

出版信息

Methods Mol Biol. 2020;2052:219-228. doi: 10.1007/978-1-4939-9748-0_13.

DOI:10.1007/978-1-4939-9748-0_13

PMID:31452165

Abstract

Cryptosporidium parvum can be reliably genetically manipulated using CRISPR/Cas9-driven homologous repair coupled to in vivo propagation within immunodeficient mice. Recent modifications have simplified the initial protocol significantly. This chapter will guide through procedures for excystation, transfection, infection, collection, and purification of transgenic Cryptosporidium parvum.

摘要

利用CRISPR/Cas9驱动的同源修复并结合免疫缺陷小鼠体内繁殖，可对微小隐孢子虫进行可靠的基因操作。最近的改进显著简化了初始方案。本章将指导微小隐孢子虫的脱囊、转染、感染、收集和纯化程序。