Mouse transgenesis in a single locus with independent regulation for multiple fluorophores.
A major barrier to complex experimental design in mouse genetics is the allele problem: combining three or more alleles is time-consuming and inefficient. Here, we solve this problem for transgenic animals with a simple modification of existing BAC transgenesis protocols, and generate triple-colored 'prism' mice in which the major cell types of the brain: neurons, astrocytes, and oligodendrocytes, are each labeled with a distinct fluorophore. All three fluorophores are expressed from the same locus, yet each fluorophore is expressed in an independent temporal and spatial pattern. All three transgenes are generally co-inherited across multiple generations with stable genomic copy number and expression patterns. This generic solution should permit more sophisticated experimental manipulations to assess functional interactions amongst populations of cell types in vivo in a more rapid and efficient manner.
Laboratory of Molecular Biology, The Rockefeller University, New York, New York, United States of America. jdougherty@genetics.wustl.edu
Joseph D
JD
Dougherty
Juliet
J
Zhang
Huifen
H
Feng
Shiaoching
S
Gong
Nathaniel
N
Heintz
D000818:Animals; D001522:Behavior, Animal; D001921:Brain / Q000166:cytology / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes / Q000378:metabolism*; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D056426:Genetic Loci / Q000235:genetics*; D040582:Inheritance Patterns / Q000235:genetics; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D050825:Synaptosomal-Associated Protein 25 / Q000378:metabolism; D019076:Transgenes / Q000235:genetics
D005456:Fluorescent Dyes; D008164:Luminescent Proteins; C494209:Snap25 protein, mouse; D050825:Synaptosomal-Associated Protein 25