Assessment of escape from X chromosome inactivation and gene expression in single human immune cells

Sven Hendrik Hagen; Jana Hennesen; Marcus Altfeld

doi:10.1016/j.xpro.2021.100641

Assessment of escape from X chromosome inactivation and gene expression in single human immune cells

STAR Protoc. 2021 Jul 20;2(3):100641. doi: 10.1016/j.xpro.2021.100641. eCollection 2021 Sep 17.

Authors

Sven Hendrik Hagen¹, Jana Hennesen², Marcus Altfeld¹

Affiliations

¹ Research Department Virus Immunology, Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Martinistrasse 52, Hamburg 20251, Germany.
² Technology Platform Flow Cytometry / FACS, Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Martinistrasse 52, Hamburg 20251, Germany.

Abstract

X-chromosomal genes escaping from X chromosome inactivation (XCI) in immune cells can contribute to sex-specific differences in immune responses. This protocol describes the specific steps to determine escape from XCI and to simultaneously quantify mRNA expression of multiple genes at the single immune cell level using a single-nucleotide polymorphism approach. The protocol furthermore allows the analysis of allele-specific expression of X-chromosomal genes. For complete details on the use and execution of this protocol, please refer to Hagen et al. (2020).

Keywords: Flow Cytometry/Mass Cytometry; Gene Expression; Immunology; Molecular Biology.

Publication types

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

MeSH terms

Alleles
Cohort Studies
DNA Primers
Flow Cytometry
Gene Expression*
Humans
Leukocytes, Mononuclear / metabolism*
Polymorphism, Single Nucleotide
Single-Cell Analysis / methods
X Chromosome Inactivation*

Substances

DNA Primers