<i>piggyBac</i>- and phiC31 integrase-mediated transgenesis in <i>Drosophila prolongata</i>

Abstract

18The development of transgenesis systems in non-model organisms provides a 19 powerful tool for molecular analysis and contributes to the understanding of phenomena

Multiplexed drug-based selection and counterselection genetic manipulations in Drosophila

“…The vector toolkit also contains all of the basic GB2.0 cloning vectors used in this work needed to adopt this cloning method. Because many of our vectors contain attB sites for ΦC31-integrase-mediated transgenesis, they can readily be transferred to additional insects, including other Drosophila and mosquito species ( Holtzman et al, 2010 ; Labbé et al, 2010 ; Meredith et al, 2013 ; Pondeville et al, 2014 ; Volohonsky et al, 2015 ; Kudo et al, 2018 ), as well as vertebrates, such as fish and mice ( Tasic et al, 2011 ; Kirchmaier et al, 2013 ; Mosimann et al, 2013 ; Roberts et al, 2014 ), that already have attP integration sites available.…”

“…The large library of DNA elements makes it possible for other labs to quickly build their own constructs and tools based on their needs. As all the markers are wholly exogenous to insects, selection and counterselection genetics can be readily applied to other drosophilids ( Holtzman et al, 2010 ; Kudo et al, 2018 ) and other insect species like mosquitoes ( Labbé et al, 2010 ; Meredith et al, 2013 ; Pondeville et al, 2014 ; Volohonsky et al, 2015 ; Matthews and Vosshall, 2020 ), provided careful selection of well-expressing promoters to ensure effective drug resistance and sensitivity. Obviously, the use of these markers in other species will have to be adapted to the particularities of their life cycles, life stages, and culture conditions.…”

Contribution of visual stimuli to mating and fighting behaviors of Drosophila prolongata

“…Microinjection was carried out in accordance with our previous study (Kudo et al . 2017). Briefly, eggs were collected for 1 h and dechorionated before injection with Femtotips II (Eppendorf, Hamburg, Germany) on an inverted phase contrast microscope (Primo vert; Carl Zeiss Microscopy, Jena, Germany).…”

“…This starvation treatment increases female receptivity and male–male aggressiveness (Kudo et al . 2017; Ando et al . 2020).…”

“…The injection solution consisted of 200 ng/μL of crRNA and trRNA, respectively, and 1,500 ng/μL of Cas9 protein (Cas9 Nuclease protein NLS; NIPPON GENE, Tokyo, Japan) in water. Microinjection was carried out in accordance with our previous study (Kudo et al 2017). Briefly, eggs were collected for 1 h and dechorionated before injection with Femtotips II (Eppendorf, Hamburg, Germany) on an inverted phase contrast microscope (Primo vert; Carl Zeiss Microscopy, Jena, Germany).…”