DOI: 10.5061/dryad.x0k6djhw3
title: Abiotic environments can modify the penetrance of transgene-based lethality systems for insect population control

description: Modern genetic biocontrol techniques for insect pest management, when
 compared to chemical insecticide spraying, offer high species specificity
 and reduced environmental impact. However, some of these methods require
 the environmental release of genetically modified insects. Because
 organisms exposed to different environments often show variability in
 phenotype and gene expression, it is likely that genetically modified
 insects will also experience environmentally mediated variation,
 potentially compromising pest control efficiency. This study examines the
 impact of temperature and nutrition on the early embryonic Tet-off
 conditional lethality system in Drosophila melanogaster. By independently
 manipulating parental and offspring environments, we assessed how
 treatment exposure influenced the probability of larval hatching and the
 transcript abundance of the transgenic system. Our findings revealed that
 (1) transgene performance distinctly responds to temperature and
 nutrition, (2) thermal stress has a greater impact when embryos, rather
 than parents, are exposed, and (3) extreme nutritional conditions can
 markedly reduce the penetrance of transgenic lethality. Although changes
 in transgene transcript abundance were observed, they did not fully
 explain the phenotypic variation, suggesting that factors downstream of
 transcription likely drive variation in transgenic lethality.
description: There are two kinds of experiments, larvae hatching and
 transcript abundance.  
 Larvae hatching - 3 files  
 Describes the frequency of hatching events in a sample of
 embryos.  
 Files:
 AbioticStress-dataset-hatching-temperature.csv
 AbioticStress-dataset-hatching-nutrition.csv
 AbioticStress-dataset-hatching-nutrition.csv
   Transcript abundance - 5
 files   Describes the cycle
 threshold (CT) values of quantitative PCR experiments
   Files:
 TransgenicExpression-dataset-nutrition-adult.csv
 TransgenicExpression-dataset-nutrition-development.csv TransgenicExpression-dataset-temperature-adult.csv TransgenicExpression-dataset-temperature-devo.csv TransgenicExpression-dataset-temperature-embryo.csv TransgenicExpression-dataset-temperature-shock.csv   This data is associated with manuscript RSPB-2025-0307, Fernan Rodrigo Perez Galvez, February 1, 2025.
description: # Abiotic environments can modify the penetrance of transgene-based
 lethality systems for insect population control
 [https://doi.org/10.5061/dryad.x0k6djhw3](https://doi.org/10.5061/dryad.x0k6djhw3) ## Description of the data and file structure Two kinds of experiments were conducted, the larvae hatching datasets describe the number of fly embryos that hatch in a sample and the transcript abundance datasets describe the cycle thresholds (CTs) of the genes of interest (driver = "TTA", effector = "HIDA5", and reference = "RP49") in transgenic embryos during the activation of the transgene under study (see manuscript).  Samples were collected in a series of blocks with multiple treatments in each of them. The experimental variables associated to the replication design and are for random effects in the statistical analysis are also included in each case. ### Files and variables #### File: AbioticStress-dataset-hatching-nutrition.csv **Description:** Describes the frequency of hatching events in a sample of embryos whose parents were exposed to nutritional treatments ##### Variables * treatment: value indicates the nutritional treatment, "1h" = high P:C ratio, "2e" = equal P:C ratio (see note 1), "3l" = low P:C ratio, "4q" = quarter diet, "5d" = desiccation treatment * stage: indicates the life stage of exposure, "1d" = development, "2a" = adult * level: verbose variable equivalent to treatment, "1high P:C" for "1h", "2equal P:C" for "2e", "3low P:C" for "3l", "4quarter" for "4q", "5diss" for "5d" * cross: identifier variable for control "c" or experimental "X" sample * replicate: indicates the replicate number of the sample, "1", "2" and "3" were used for analysis, samples with value "0" were considered burnout samples and were not included * block: indicates the block number that the sample belongs, variable used for modeling random effects * total: total number of embryos in the sample * hatch: number of embryos hatched * hrate: hatching rate calculated as "hrate = hatch / total" * cohort: indicates the experimental replicate, that is cohort "1" for the first replicate, etc. #### File: AbioticStress-dataset-hatching-shock.csv **Description:** Describes the frequency of hatching events in a sample of embryos whose parents were exposed to thermal shock treatments ##### Variables * treatment: value indicates shock treatments as "k" * temp: treatment level, "control" = 25°C, "heat" = 34°C, "cold" = 4°C * hours: numerical variable indicating the number of hours, either "1", "2", "4", "24", or "48" * cross: identifier variable for control "0" or experimental "1" sample * replicate: indicates the replicate number of the experiment, "1", "2" and "3" * total: total number of embryos in the sample * hatch: number of embryos hatched * hrate: hatching rate calculated as "hrate = hatch / total" #### File: TransgenicExpression-dataset-nutrition-adult.csv **Description:** Describes the cycle threshold (CT) values of quantitative PCR experiments in embryos whose parents were exposed to nutritional treatments during adulthood ##### Variables * Experiment: identifier and metadata of file * CT: cycle threshold * Well Position: well position in the 96-well plate * Gene: target gene, reference = "RP49", effector = "HIDA5", driver = "TTA" * Level: nutritional treatment, "High" for High P:C, "Equal" for Equal P:C, "Low" for Low P:C, "Qrtr" for Quarter caloric restriction diet, "Diss" for desiccation treatment. * ReplicateB: biological replicate * ReplicateT: technical replicate #### File: AbioticStress-dataset-hatching-temperature.csv **Description:** Describes the frequency of hatching events in a sample of embryos whose parents were exposed to thermal treatments during adulthood ##### Variables * treatment: value "t" indicates temperature treatments - stage: indicates the life stage of exposure, "1d" = development, "2a" = adult, "3e" = embryo - level: treatment level ID with joint temperature and "C" for degree Celsius - temp: treatment temperature - cross: identifier variable for control "c" or experimental "X" sample - replicate: indicates the replicate number of the sample, "1", "2" and "3" were used for analysis, samples with value "0" were considered burnout samples and were not included in the analysis - block: indicates the block number that the sample belongs, variable used for modeling random effects - total: total number of embryos in the sample - hatch: number of embryos hatched - hrate: hatching rate calculated as "hrate = hatch / total" - cohort: indicates the experimental replicate, that is cohort "1" for the first replicate, etc. #### File: TransgenicExpression-dataset-temperature-adult.csv **Description:** Describes the cycle threshold (CT) values of quantitative PCR experiments in embryos whose parents were exposed to thermal treatments during adulthood ##### Variables * Experiment: identifier and metadata of file * CT: cycle threshold * Well Position: well position in the 96-well plate * Gene: target gene, reference = "RP49", effector = "HIDA5", driver = "TTA" * Level: treatment temperature, either 19, 22, 25, 28, or 31 degrees Celsius * ReplicateB: biological replicate * ReplicateT: technical replicate NOTE: Missing data for tTA reading (rows 170-178), interestingly the same biological samples indeed provided information on the other qPCR targets. We interpret this as "undetected". #### File: TransgenicExpression-dataset-nutrition-development.csv **Description:** Describes the cycle threshold (CT) values of quantitative PCR experiments in embryos whose mothers were exposed to nutritional treatments during development ##### Variables * Experiment: identifier and metadata of file * CT: cycle threshold * Well Position: well position in the 96-well plate * Gene: target gene, reference = "RP49", effector = "HIDA5", driver = "TTA" * Level: nutritional treatment, "High" for High P:C, "Equal" for Equal P:C, "Low" for Low P:C, "Quarter" for Quarter caloric restriction diet * ReplicateB: biological replicate * ReplicateT: technical replicate NOTE: Biological and technical replicates (3x) from rows 11, 12, 13, 23, 24, 25, 59, 60, 61, 71, 72, 73, 107, 108, 109, 119, 120, 121 were removed from dataset for quality reasons. Their impact in the experimental design was the reduction of one biological replicate per treatment, leaving at least 3 biological replicates with 3 technical replicates each. #### File: TransgenicExpression-dataset-temperature-devo.csv **Description:** Describes the cycle threshold (CT) values of quantitative PCR experiments in embryos whose mothers were exposed to thermal treatments during development ##### Variables * Experiment: identifier and metadata of file * CT: cycle threshold * Well Position: well position in the 96-well plate * Gene: target gene, reference = "RP49", effector = "HIDA5", driver = "TTA" * Level: treatment temperature, either 19, 22, 25, or 28 degrees Celsius * ReplicateB: biological replicate * ReplicateT: technical replicate #### File: TransgenicExpression-dataset-temperature-embryo.csv **Description:** Describes the cycle threshold (CT) values of quantitative PCR experiments in embryos exposed to thermal treatments ##### Variables * Experiment: identifier and metadata of file * CT: cycle threshold * Well Position: well position in the 96-well plate * Gene: target gene, reference = "RP49", effector = "HIDA5", driver = "TTA" * Level: treatment temperature, either 16,19, 22, 25, 28 or 31 degrees Celsius * ReplicateB: biological replicate * ReplicateT: technical replicate #### File: TransgenicExpression-dataset-temperature-shock.csv **Description:** Describes the cycle threshold (CT) values of quantitative PCR experiments in embryos whose parent were exposed to thermal shock ##### Variables * Experiment: identifier and metadata of file * CT: cycle threshold * Well Position: well position in the 96-well plate * Gene: target gene, reference = "RP49", effector = "HIDA5", driver = "TTA" * Level: treatment level, "Cold" = cold shock, "No" = control, "Heat" = heat shock * Time: (only shock file) - hours after shock * ReplicateB: biological replicate * ReplicateT: technical replicate NOTE: Biological replicate from rows 170, 171, 172 were removed from dataset for quality reasons