standard Rational Design of Transgenic Experiments

Transposon

Cut-and-paste transposons like piggyBac make good gene vectors although their performance is highly variable as the authors show in their paper.

Gregory et al (2016) have published a very interesting meta analysis of piggyBac transformation data in Insect Molecular Biology that is worth taking a look at if you are thinking about making transgenic insects.

The focus of the study is the efficiency of insect transformation using gene vectors constructed from the piggyBac transposon.   The authors use published data as well as unpublished data from Oxitec, Ltd.  Oxitec has developed a conditional lethal population suppression strategy that depends upon the introduction of their custom transgene into the germline of target species using piggyBac-based vectors.  They have been working with a number of insect species and accumulated data regarding their success relative to the number of injections, survival post injection, transformation frequencies etc.

AppScreenShot

Screen shot of the Goldilocks App for planning piggyBac transformation experiments in insects.

The authors provide some interesting and enlightening summary statistics on transformation efficiencies by species and by insect order.  There is lots of variance as expected but a look at those data can lead you to begin to have sober thoughts about what you are about to attempt (and that is one of the points of the analysis).  Most of the transformation efficiency data are between 1 and 10 % – not great, but workable.  Efficiencies in Lepidoptera are lower than those in Diptera.

Size seems to matter, as those who have worked with transposon vectors are aware.  This was first reported many years ago by Alan Spradling based on P element data. Again, lots of variance but there is a negative trend.

Goldy

The Goldilocks model helps investigators get their effort just right based on survival and transformation efficiency.

The authors have created an application – The Goldilocks Decision Model – that is intended to help researchers decide the level of effort they might need to consider given certain parameters that the user can manipulate like survival, transformation efficiency.  This is interesting and certainly gives investigators some sense of how they might design their experiments or adjust their expectations if they are paying for someone else to do it for you.

This paper is worth taking a good look at, as is their model, which can be found at https://mammykins.shinyapps.io/App-gold

 

Gregory, M., Alphey, L., Morrison, N. I., Shimeld, S. M., 2016 Insect transformation with piggyBac: getting the number of injections just right. Insect Mol Biol: n/a-n/adoi: 10.1111/imb.12220.

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