清脆的
基因
生物
抄写(语言学)
转录调控
激活剂(遗传学)
转基因
CRISPR干扰
遗传学
计算生物学
转录因子
细胞生物学
Cas9
语言学
哲学
作者
Yuting Han,Xinyi Lu,Yutong Li,Yuhao Qiu,Xizhi Dong,Xiaochen Li,Xu Si,Qingfei Liu,Jian-Quan Ni
出处
期刊:Methods in molecular biology
日期:2022-01-01
卷期号:: 177-199
标识
DOI:10.1007/978-1-0716-2541-5_8
摘要
Overexpression is one of the classical approaches to study pleiotropic functions of genes of interest. To achieve overexpression, we often increase the transcription by introducing genes on exogenous vectors or by using the CRISPR/dCas9-based transcriptional activation system. To date, the most efficient CRISPR/dCas9-based transcriptional activator is the Synergistic Activation Mediator (SAM) system whereby three different transcriptional activation domains are directly fused to dCas9 and MS2 phage Coat Protein (MCP), respectively, and the system in Drosophila is named flySAM. Here we describe the effective and convenient transcriptional activation system, flySAM, starting from vector construction, microinjection, and transgenic fly selection to the phenotypic analysis.
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