通过CRISPR-Cas9基因编辑,生成携带B细胞受限单个或多重功能丧失突变的小鼠模型
Elisa Ten Hacken1, Michaela Gruber2, María Hernández-Sánchez3
1Department of Medical Oncology, Dana Farber Cancer Institute, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.
STAR protocols
|September 20, 2023
概括
这项研究详细介绍了一项协议,用于通过工程干细胞来创建淋巴增殖性疾病的小鼠模型,用于B细胞特异性基因编辑. 这些模型有助于研究这些血液癌症的复杂遗传驱动因素.
科学领域:
- 免疫学 免疫学 免疫学
- 遗传学 是一个遗传学.
- 血液学 血液学 血液学
背景情况:
- 淋巴增殖性疾病是一组由淋巴细胞异常增殖为特征的疾病.
- 了解这些疾病的遗传驱动因素对于开发有效治疗方法至关重要.
- 现有的小鼠模型可能无法完全回顾人类淋巴增殖性疾病的复杂性.
研究的目的:
- 开发一个协议,用于生成B细胞受限的小鼠模型的淋巴增殖性疾病.
- 为了研究这些疾病典型的功能丧失遗传驱动因素.
- 为模拟复杂疾病特征提供一个多功能平台.
主要方法:
- 具有B细胞受限Cas9表达的小鼠菌株的干细胞工程.
- 一种表达 sgRNA-mCherry 的晶状病毒制剂.
- 隔离和体外转化造血干细胞和祖细胞.
- 将工程细胞移植到接受者小鼠体内,并验证基因编辑.
主要成果:
- 通过使用描述的协议,成功生成了B细胞受限小鼠模型.
- 在干细胞工程之后,在B细胞中展示了高效的基因编辑.
- 验证小鼠模型作为研究淋巴增殖性疾病的平台.
结论:
- 本方案提供了一种强大的方法,用于为淋巴增殖性疾病创建基因工程小鼠模型.
- 这些模型有助于研究特定的遗传驱动因素和疾病机制.
- 工程小鼠是促进研究B细胞恶性瘤和相关疾病的宝贵工具.
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