通过敲除将CRISPR-Cas9电穿孔导入恶性B细胞以进行靶基因功能丧失研究。

Electroporation of CRISPR-Cas9 into Malignant B Cells for Loss-of-Function Studies of Target Gene Via Knockout.

作者信息

Yu Jian

机构信息

Institute of Nanotechnology for Single Cell Analysis (INSCA), Beijing Advanced Innovation Center for Biomedical Engineering, Beihang University, Beijing, China.

Beijing Advanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, Beihang University, Beijing, China.

出版信息

Methods Mol Biol. 2020;2050:85-90. doi: 10.1007/978-1-4939-9740-4_9.

DOI:10.1007/978-1-4939-9740-4_9

PMID:31468482

Abstract

CRISPR-Cas9 is a unique technology that enables geneticists and medical researchers to edit genomic DNA for studying biology, pathogenesis, and molecular basis of treatment in malignant B cells. Unfortunately, malignant B cells are extremely difficult to transfect by most traditional methods. In this chapter, we describe the use of the Nucleofector™ Technology-based electroporation system with optimized transfection conditions for generating a malignant B cell model, JEKO-1, with ROR1-gene knockout via CRISPR-Cas9 technology.

摘要

CRISPR-Cas9是一项独特的技术，它使遗传学家和医学研究人员能够编辑基因组DNA，以研究恶性B细胞的生物学、发病机制和治疗的分子基础。不幸的是，大多数传统方法极难转染恶性B细胞。在本章中，我们描述了基于Nucleofector™技术的电穿孔系统的使用，该系统具有优化的转染条件，用于通过CRISPR-Cas9技术生成ROR1基因敲除的恶性B细胞模型JEKO-1。