Selecting Transgenic Mosquito Larvae with Puromycin

The selection of mosquito transgenic larvae using a nonfluorescent approach can be advantageous to reserve fluorophores for downstream applications, such as immunostaining or for the study of promoter activity by cloning a fluorescence reporter gene under the control of that promoter. We previously...

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Bibliographic Details
Published inCold Spring Harbor protocols Vol. 2024; no. 6; p. pdb.prot108308
Main Author Marois, Eric
Format Journal Article
LanguageEnglish
Published United States 03.06.2024
Subjects
Animals
Animals, Genetically Modified - genetics
Anopheles - genetics
Larva - genetics
Puromycin - pharmacology
Selection, Genetic
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Summary:The selection of mosquito transgenic larvae using a nonfluorescent approach can be advantageous to reserve fluorophores for downstream applications, such as immunostaining or for the study of promoter activity by cloning a fluorescence reporter gene under the control of that promoter. We previously reported that puromycin selection is efficient in transgenic or larvae expressing an selection marker. A concentration of puromycin of >10 µg/mL is lethal for larvae, unless they carry the resistance gene, conferring them resistance to puromycin concentrations of 25-80 µg/mL. A drawback of this fully dominant selection marker is that, unlike with fluorescence markers, homozygous transgenics cannot be distinguished from heterozygotes. Here, we outline the procedure for selecting puromycin-resistant transgenic larvae.
ISSN:1559-6095
DOI:10.1101/pdb.prot108308