A knock-in allele of En1 expressing dre recombinase.

Abstract

Engrailed 1 (En1) is a homeobox-containing transcription factor expressed during development in diverse tissues, including the embryonic midbrain and anterior hindbrain. To facilitate investigation of genetic and developmental heterogeneity among cells with a history of En1 expression, we have generated En1(Dre) , a knock-in allele expressing Dre recombinase. En1(Dre) can be used with existing Cre and Flp recombinase lines for genetic intersectional labeling, fate mapping, and functional manipulation of subpopulations of cells characterized by transient expression of En1. To avoid disrupting En1 function, the Dre cDNA is inserted at the 3' end of the En1 coding sequence, together with a viral 2A peptide to mediate translation of separate EN1 and Dre proteins. Consequently, viable and fertile En1(Dre) homozygotes can be used to increase the proportion of useful genotypes produced in complex crosses. The pattern of Dre expression from En1(Dre) is indistinguishable from wild-type En1 expression in mid-gestation mouse embryos, and En1(Dre) controls Dre-responsive indicator alleles by efficiently recombining rox sites in vivo. Through the application of genetic tools that allow manipulation of cells based on combinatorial expression of multiple distinct recombinases, En1(Dre) will significantly extend the ability to target important subpopulations of neurons and other cells within the broader En1 expression domain. genesis 54:447-454, 2016. Published 2016. This article is a US Government work and is in the public domain in the USA.