A new genetic tool to modify and understand gene function

A new genetic tool to modify and understand gene function
Single cells expressing the iSuRe-Cre allele (in yellow) and the conventional reporter (in green). Only the yellow cell has the gene deleted. Credit: CNIC

Scientists of the National Center for Cardiovascular Research (CNIC) led by Rui Benedito have developed a new genetic tool called iSuRe-Cre that provides certainty in Cre-inducible genetic modifications, a key technique for understanding gene function.

Most analysis of gene in biomedical research relies on the use of Cre-lox technology. Since its introduction in 1994, this technology has revolutionized because it allows scientists to eliminate or activate the function of any gene in any cell type in the mouse. Using this technology, it is possible to investigate the precise role of almost any mouse gene in any cell type, and at a defined time point, all of which are crucial requirements for understanding the function of genes during organ development, physiology and disease.

Cre-Lox technology allows the regulation of gene expression at any time or in any cell type thanks to the ability of the Cre recombinase protein to recognize and recombine lox sites introduced at specific locations in the mouse genome, leading to the deletion of the genes being studied.

Despite the major impact of Cre-loxP technology on biomedical research, numerous studies have demonstrated the need for caution in its use. The main problem is that often, the Cre activity level is insufficient to fully recombine and eliminate expression of the target gene, generating uncertainty about whether the desired has been achieved.

Because the Cre-loxP recombination event is invisible, genetic and fluorescent reporters have been developed to track Cre activity so that cells reaching a certain threshold of Cre activity are labeled. These reporters have become established as essential genetic tools for monitoring any conditional genetic study. However, as study author Macarena Fernández-Chacón points out, many studies have shown that Cre-induced reporter expression can occur even though other genes with loxP sites are not completely eliminated.

To overcome this technical hurdle, the CNIC team have developed an innovative method based on a new allele called iSuRe-Cre. iSuRe-Cre is compatible with all existing Cre/CreERT2/lox alleles and guarantees high Cre activity in the cells that express the fluorescent reporter. This ultimately increases the efficiency and reliability of the analysis of Cre-dependent gene function. Moreover, the use of the new iSuRe-Cre mice permits the induction of multiple genetic deletions in the same cell. This important property allows the study of functional genetic interactions or epistasis, or in other words, how the function of one or more genes depends on the function of another.

Macarena Fernández-Chacón says, "The use of this new genetic tool will significantly change research projects that depend on the Cre-lox system, because now we can see and be confident about the location of that have one or more eliminated."

Lead investigator Rui Benedito adds that the new genetic tool will be of great interest in biomedical research "because it significantly increases the ease, efficiency, and reliability of genetic modification in the mouse, the most widely used animal model in research." The new study is published in Nature Communications.


Explore further

Scientists develop new methods for analyzing gene function

Journal information: Nature Communications

Provided by Centro Nacional de Investigaciones Cardiovasculares Carlos III (F.S.P.)
Citation: A new genetic tool to modify and understand gene function (2019, May 22) retrieved 19 June 2019 from https://medicalxpress.com/news/2019-05-genetic-tool-gene-function.html
This document is subject to copyright. Apart from any fair dealing for the purpose of private study or research, no part may be reproduced without the written permission. The content is provided for information purposes only.
8 shares

Feedback to editors

User comments

Please sign in to add a comment. Registration is free, and takes less than a minute. Read more