Affordable Access

deepdyve-link
Publisher Website

Poor Concordance of Floxed Sequence Recombination in Single Neural Stem Cells: Implications for Cell Autonomous Studies.

Authors
  • Dause, Tyler Joseph1
  • Kirby, Elizabeth Diana2, 3, 4
  • 1 Deptartment of Psychology, The Ohio State University, Columbus, OH 43210.
  • 2 Deptartment of Psychology, The Ohio State University, Columbus, OH 43210 [email protected]
  • 3 Deptartment of Neuroscience, The Ohio State University, Columbus, OH 43210.
  • 4 The Chronic Brain Injury Program, The Ohio State University, Columbus, OH 43210.
Type
Published Article
Journal
eneuro
Publisher
Society for Neuroscience
Publication Date
Jan 01, 2020
Volume
7
Issue
2
Identifiers
DOI: 10.1523/ENEURO.0470-19.2020
PMID: 32079584
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

To manipulate target gene function in specific adult cell populations, tamoxifen (TAM)-dependent CreERT2 is widely used to drive inducible, site-specific recombination of loxP flanked sequences. In studies of cell autonomous target gene function, it is common practice to combine these CreERT2-lox systems with a ubiquitously expressed stop-floxed fluorescent reporter gene to identify single cells supposedly undergoing target gene recombination. Here, we studied the reliability of using Cre-induced recombination of one gene to predict recombination in another gene at the single-cell level in adult hippocampal neural stem and progenitor cells (NSPCs). Using both probabilistic predictions in a generic experimental paradigm, as well as a mouse model with two separate stop-floxed reporters plus a Nestin promoter-driven CreERT2, we found that, in individual cells, recombination of one gene was a poor predictor of recombination in another. This poor concordance in floxed sequence recombination across genes suggests that use of stop-floxed reporters to investigate cell autonomous gene function may not be universally reliable and could lead to false conclusions. Copyright © 2020 Dause and Kirby.

Report this publication

Statistics

Seen <100 times