A protocol for CRISPR-mediated activation and repression of human endogenous retroviruses in human pluripotent stem cells

Vidya Padmanabhan Nair; Jens Mayer; Michelle Vincendeau

doi:10.1016/j.xpro.2022.101281

A protocol for CRISPR-mediated activation and repression of human endogenous retroviruses in human pluripotent stem cells

STAR Protoc. 2022 Apr 12;3(2):101281. doi: 10.1016/j.xpro.2022.101281. eCollection 2022 Jun 17.

Authors

Vidya Padmanabhan Nair¹, Jens Mayer², Michelle Vincendeau¹

Affiliations

¹ Institute of Virology, Helmholtz Zentrum München, 85764 Neuherberg, Germany.
² Institute of Human Genetics, Medical Faculty, University of Saarland, 66421 Homburg, Germany.

Abstract

Human endogenous retroviruses (HERVs) comprise many regulatory elements and can regulate host gene activity at different expression levels via multiple mechanisms. Here, we introduce a step-by-step protocol to activate or repress transcription of HERV-K(HML-2) elements using the CRISPRa and CRISPRi technologies in human embryonic stem cells. This protocol can help deciphering the functional role of HERV-K(HML-2) elements in critical biological processes. The protocol may easily be adapted to other cell lines and HERV groups with relatively low sequence heterogeneity. For complete details on the use and execution of this protocol, please refer to Padmanabhan Nair et al. (2021).

Keywords: CRISPR; Developmental biology.

Publication types

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

MeSH terms

Clustered Regularly Interspaced Short Palindromic Repeats / genetics
Endogenous Retroviruses* / genetics
Humans
Pluripotent Stem Cells*