How to Design a Genetic Mating Scheme: A Basic Training Package for Drosophila Genetics

Abstract

Drosophila melanogaster is a powerful model organism for biological research. The essential and common instrument of fly research is genetics, the art of applying Mendelian rules in the specific context of Drosophila with its unique classical genetic tools and the breadth of modern genetic tools and strategies brought in by molecular biology, transgenic technologies and the use of recombinases. Training newcomers to fly genetics is a complex and time-consuming task but too important to be left to chance. Surprisingly, suitable training resources for beginners currently are not available. Here we provide a training package for basic Drosophila genetics, designed to ensure that basic knowledge on all key areas is covered while reducing the time invested by trainers. First, a manual introduces to fly history, rationale for mating schemes, fly handling, Mendelian rules in fly, markers and balancers, mating scheme design, and transgenic technologies. Its self-study is followed by a practical training session on gender and marker selection, introducing real flies under the dissecting microscope. Next, through self-study of a PowerPoint presentation, trainees are guided step-by-step through a mating scheme. Finally, to consolidate knowledge, trainees are asked to design similar mating schemes reflecting routine tasks in a fly laboratory. This exercise requires individual feedback but also provides unique opportunities for trainers to spot weaknesses and strengths of each trainee and take remedial action. This training package is being successfully applied at the Manchester fly facility and may serve as a model for further training resources covering other aspects of fly research.

Neighboring genes for DNA-binding proteins rescue male sterility in Drosophila hybrids

“…Injected embryos were subsequently raised in the laboratory, and emerging individual virgin adult w − flies were backcrossed to SimB. Progeny were screened via presence or absence of the w + eye marker (74), and those containing the pB[w + ] transposed DNA insert were selected to establish stable male transgenic lines. Altogether, the progeny of 429 (agt) and 160 (Taf1) injected adult flies was scored, all constructs considered, and yielded similar transformation efficiencies of 2.79% and 3.1%.…”

Using fly genetics to dissect the cytoskeletal machinery of neurons during axonal growth and maintenance

“…Drosophila as a model to study cytoskeletal machinery during axonal growth One organism in which combinatorial genetics has been and continues to be most successfully applied to the study of complex biological problems is the fruit fly Drosophila melanogaster (Keller, 1996;Roote and Prokop, 2013). Work in Drosophila is low cost, impressively fast, and particularly amenable to genetic manipulations and unbiased genetic screens (Sánchez-Soriano et al, 2007).…”

Functional and Genetic Analysis of Spectraplakins in Drosophila

“…Using simple classical genetic crosses allows the fast generation of animals combining multiple genetic aberrations or transgenic constructs which can then be applied for studies of spectraplakin functions in development and disease. The basic knowledge required for this work is readily accessible and has recently been put together in a comprehensive manual for newcomers to Drosophila research (Prokop, 2013b;Roote & Prokop, 2013). Also the generation of transgenic animals takes just 2 months from injection to first experimentation and can be outsourced at low cost.…”

“…1B); shot kakP2 was therefore used for structure-function analyses, complementary to transgenic rescues strategies (see Section 3) with the Shot-LC construct similarly lacking the first CH domain (Bottenberg et al, 2009;R€ oper & Brown, 2003). Another important application of shot mutant alleles or transgenic tools is to capitalize on the ease of Drosophila mating schemes (Roote & Prokop, 2013) and combine them with mutant alleles of other genes. The key principle is to use partial LOF conditions for shot (e.g., a heterozygous condition for a null allele, such as shot sf20/+ ) and for another gene, where neither of the two alone causes a phenotype.…”