Laboratory of Synthetic Genome Biology
Developing biological engineering approaches to understand epigenetics and genome organization in C. elegans

Froekjaer-Jensen et al (2008) MosSCI

Single-copy insertion of transgenes in Caenorhabditis elegans.

Froekjaer-Jensen et al. (2008). Single-copy insertion of transgenes in Caenorhabditis elegans. Nature Genetics. ​
Frøkjaer-Jensen C, Davis MW, Hopkins CE, Newman BJ, Thummel JM, Olesen SP, Grunnet M, Jorgensen EM.
Transgenesis, Endogenous expression, Germline expression, Single-copy transgenes, Jorgensen Lab, University of Utah
2008
At present, transgenes in Caenorhabditis elegans are generated by injecting DNA into the germline. The DNA assembles into a semistable extrachromosomal array composed of many copies of injected DNA. These transgenes are typically overexpressed in somatic cells and silenced in the germline. We have developed a method that inserts a single copy of a transgene into a defined site. Mobilization of a Mos1 transposon generates a double-strand break in noncoding DNA. The break is repaired by copying DNA from an extrachromosomal template into the chromosomal site. Homozygous single-copy insertions can be obtained in less than 2 weeks by injecting approximately 20 worms. We have successfully inserted transgenes as long as 9 kb and verified that single copies are inserted at the targeted site. Single-copy transgenes are expressed at endogenous levels and can be expressed in the female and male germlines.​​​