CRISPR-Based Transcriptional Activation in Drosophila

Yuting Han; Xinyi Lu; Yutong Li; Yuhao Qiu; Xizhi Dong; Xiaochen Li; Xu Si; Qingfei Liu; Jian-Quan Ni

doi:10.1007/978-1-0716-2541-5_8

CRISPR-Based Transcriptional Activation in Drosophila

Methods Mol Biol. 2022:2540:177-199. doi: 10.1007/978-1-0716-2541-5_8.

Authors

Yuting Han¹, Xinyi Lu¹, Yutong Li¹, Yuhao Qiu¹, Xizhi Dong¹, Xiaochen Li¹, Xu Si¹, Qingfei Liu¹, Jian-Quan Ni²

Affiliations

¹ Gene Regulatory Lab, School of Medicine, Tsinghua University, Beijing, China.
² Gene Regulatory Lab, School of Medicine, Tsinghua University, Beijing, China. nijq@mail.tsinghua.edu.cn.

PMID: 35980578
DOI: 10.1007/978-1-0716-2541-5_8

Abstract

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.

Keywords: CRISPR/dCas9; Drosophila; Overexpression; Transcriptional activation; flySAM.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Animals, Genetically Modified
CRISPR-Cas Systems* / genetics
Drosophila* / genetics
Drosophila* / metabolism
Transcription Factors / metabolism
Transcriptional Activation

Substances

Transcription Factors