Genetic engineering of sex chromosomes for batch cultivation of non-transgenic, sex-sorted males

The field performance of Sterile Insect Technique (SIT) is improved by sex-sorting and releasing only sterile males. This can be accomplished by resource-intensive separation of males from females by morphology. Alternatively, sex-ratio biasing genetic constructs can be used to selectively remove on...

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Published inPLoS genetics Vol. 16; no. 11; p. e1009180
Main Authors Das, Siba R., Maselko, Maciej, Upadhyay, Ambuj, Smanski, Michael J.
Format Journal Article
LanguageEnglish
Published United States Public Library of Science 02.11.2020
Public Library of Science (PLoS)
Subjects
Animal genetic engineering
Animals
Animals, Genetically Modified - physiology
Antibiotics
Biology and Life Sciences
Chromosomes
Design
Drosophila melanogaster - genetics
Engineering and Technology
Female
Females
Gene expression
Gene pool
Genetic aspects
Genetic crosses
Genetic engineering
Genetic Engineering - methods
Genetic research
Insect Control - methods
Insect sterilization
Insects
Male
Males
Medicine and Health Sciences
Methods
Models, Animal
Pest Control, Biological - methods
Pests
Phenotypes
Research and Analysis Methods
Sex chromosomes
Sex Chromosomes - genetics
Sex Determination Analysis - methods
Sex Determination Processes - genetics
Sex preselection
Sex ratio
Statistical analysis
Transgenes - genetics
X chromosomes
Australia
United States
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Abstract The field performance of Sterile Insect Technique (SIT) is improved by sex-sorting and releasing only sterile males. This can be accomplished by resource-intensive separation of males from females by morphology. Alternatively, sex-ratio biasing genetic constructs can be used to selectively remove one sex without the need for manual or automated sorting, but the resulting genetically engineered (GE) control agents would be subject to additional governmental regulation. Here we describe and demonstrate a genetic method for the batch production of non-GE males. This method could be applied to generate the heterogametic sex (XY, or WZ) in any organism with chromosomal sex determination. We observed up to 100% sex-selection with batch cultures of more than 10 3 individuals. Using a stringent transgene detection assay, we demonstrate the potential of mass production of transgene free males.
AbstractList The field performance of Sterile Insect Technique (SIT) is improved by sex-sorting and releasing only sterile males. This can be accomplished by resource-intensive separation of males from females by morphology. Alternatively, sex-ratio biasing genetic constructs can be used to selectively remove one sex without the need for manual or automated sorting, but the resulting genetically engineered (GE) control agents would be subject to additional governmental regulation. Here we describe and demonstrate a genetic method for the batch production of non-GE males. This method could be applied to generate the heterogametic sex (XY, or WZ) in any organism with chromosomal sex determination. We observed up to 100% sex-selection with batch cultures of more than 103 individuals. Using a stringent transgene detection assay, we demonstrate the potential of mass production of transgene free males.The field performance of Sterile Insect Technique (SIT) is improved by sex-sorting and releasing only sterile males. This can be accomplished by resource-intensive separation of males from females by morphology. Alternatively, sex-ratio biasing genetic constructs can be used to selectively remove one sex without the need for manual or automated sorting, but the resulting genetically engineered (GE) control agents would be subject to additional governmental regulation. Here we describe and demonstrate a genetic method for the batch production of non-GE males. This method could be applied to generate the heterogametic sex (XY, or WZ) in any organism with chromosomal sex determination. We observed up to 100% sex-selection with batch cultures of more than 103 individuals. Using a stringent transgene detection assay, we demonstrate the potential of mass production of transgene free males.
The field performance of Sterile Insect Technique (SIT) is improved by sex-sorting and releasing only sterile males. This can be accomplished by resource-intensive separation of males from females by morphology. Alternatively, sex-ratio biasing genetic constructs can be used to selectively remove one sex without the need for manual or automated sorting, but the resulting genetically engineered (GE) control agents would be subject to additional governmental regulation. Here we describe and demonstrate a genetic method for the batch production of non-GE males. This method could be applied to generate the heterogametic sex (XY, or WZ) in any organism with chromosomal sex determination. We observed up to 100% sex-selection with batch cultures of more than 10.sup.3 individuals. Using a stringent transgene detection assay, we demonstrate the potential of mass production of transgene free males.
The field performance of Sterile Insect Technique (SIT) is improved by sex-sorting and releasing only sterile males. This can be accomplished by resource-intensive separation of males from females by morphology. Alternatively, sex-ratio biasing genetic constructs can be used to selectively remove one sex without the need for manual or automated sorting, but the resulting genetically engineered (GE) control agents would be subject to additional governmental regulation. Here we describe and demonstrate a genetic method for the batch production of non-GE males. This method could be applied to generate the heterogametic sex (XY, or WZ) in any organism with chromosomal sex determination. We observed up to 100% sex-selection with batch cultures of more than 10 3 individuals. Using a stringent transgene detection assay, we demonstrate the potential of mass production of transgene free males.
Sterilized insects are released into the wild where sterile males compete with wild males to mate with wild females. Since females of many pest insects only mate once in their lifetime, mating with a sterile male prevents successful reproduction. Numbers above bars show total number of progeny produced from three biological replicates. * indicates statistically significant difference from expected 50:50 male:female sex ratio (chi-squared test, p < .05). ** indicates a statistically significant difference between the +tet and -tet groups (chi-squared test, p<0.001). https://doi.org/10.1371/journal.pgen.1009180.g002 To test the genetic circuit design, we engineered D. melanogaster due to its powerful genetic toolkit and also because its serves as a model for other insect pests. [...]both DmXL-tTA and DmYL-tTA produced a sufficiently lethal phenotype in the absence of tetracycline to remove the transgene from the accessible gene pool (Fig 2b and 2c). Sub-stoichiometric ratio of mixed-sex DmXLtTA to female DmYLtTA sufficient for non-transgenic male production Non-transgenic males can be generated by crossing non-transgenic females produced by the DmYL-tTA strain and males from a mixed-sex true-breeding population of DmXL-tTA flies (Fig 1c).
The field performance of Sterile Insect Technique (SIT) is improved by sex-sorting and releasing only sterile males. This can be accomplished by resource-intensive separation of males from females by morphology. Alternatively, sex-ratio biasing genetic constructs can be used to selectively remove one sex without the need for manual or automated sorting, but the resulting genetically engineered (GE) control agents would be subject to additional governmental regulation. Here we describe and demonstrate a genetic method for the batch production of non-GE males. This method could be applied to generate the heterogametic sex (XY, or WZ) in any organism with chromosomal sex determination. We observed up to 100% sex-selection with batch cultures of more than 103 individuals. Using a stringent transgene detection assay, we demonstrate the potential of mass production of transgene free males.
The field performance of Sterile Insect Technique (SIT) is improved by sex-sorting and releasing only sterile males. This can be accomplished by resource-intensive separation of males from females by morphology. Alternatively, sex-ratio biasing genetic constructs can be used to selectively remove one sex without the need for manual or automated sorting, but the resulting genetically engineered (GE) control agents would be subject to additional governmental regulation. Here we describe and demonstrate a genetic method for the batch production of non-GE males. This method could be applied to generate the heterogametic sex (XY, or WZ) in any organism with chromosomal sex determination. We observed up to 100% sex-selection with batch cultures of more than 10 3 individuals. Using a stringent transgene detection assay, we demonstrate the potential of mass production of transgene free males. In this manuscript, we describe and demonstrate in a model system a new genetic engineering approach for producing sex-sorted, non-transgenic batches of an organism with genetic sex determination. This approach could impact pest control programs using Sterile Insect Technique.
Sterilized insects are released into the wild where sterile males compete with wild males to mate with wild females. Since females of many pest insects only mate once in their lifetime, mating with a sterile male prevents successful reproduction. Numbers above bars show total number of progeny produced from three biological replicates. * indicates statistically significant difference from expected 50:50 male:female sex ratio (chi-squared test, p < .05). ** indicates a statistically significant difference between the +tet and -tet groups (chi-squared test, p<0.001). https://doi.org/10.1371/journal.pgen.1009180.g002 To test the genetic circuit design, we engineered D. melanogaster due to its powerful genetic toolkit and also because its serves as a model for other insect pests. [...]both DmXL-tTA and DmYL-tTA produced a sufficiently lethal phenotype in the absence of tetracycline to remove the transgene from the accessible gene pool (Fig 2b and 2c). Sub-stoichiometric ratio of mixed-sex DmXLtTA to female DmYLtTA sufficient for non-transgenic male production Non-transgenic males can be generated by crossing non-transgenic females produced by the DmYL-tTA strain and males from a mixed-sex true-breeding population of DmXL-tTA flies (Fig 1c).
Audience Academic
Author Smanski, Michael J.
Das, Siba R.
Upadhyay, Ambuj
Maselko, Maciej
AuthorAffiliation 2 Biotechnology Institute, University of Minnesota, Saint Paul, MN, United States of America
HudsonAlpha Institute for Biotechnology, UNITED STATES
1 Department of Biochemistry, Molecular Biology, and Biophysics, Saint Paul, MN, United States of America
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BackLink https://www.ncbi.nlm.nih.gov/pubmed/33137115$$D View this record in MEDLINE/PubMed
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CitedBy_id crossref_primary_10_3390_insects12030243
crossref_primary_10_7554_eLife_71230
crossref_primary_10_1111_imb_12836
crossref_primary_10_1073_pnas_2107413118
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Authors SD, MM, and MS are co-founders of Novoclade, LLC, and have filed patent application PCT/US2020/053749 related to this manuscript.
Current address: Applied BioSciences. Macquarie University, Sydney NSW, Australia. CSIRO Synthetic Biology Future Science Platform, Sydney NSW, Australia
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Snippet The field performance of Sterile Insect Technique (SIT) is improved by sex-sorting and releasing only sterile males. This can be accomplished by...
Sterilized insects are released into the wild where sterile males compete with wild males to mate with wild females. Since females of many pest insects only...
Sterilized insects are released into the wild where sterile males compete with wild males to mate with wild females. Since females of many pest insects only...
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StartPage e1009180
SubjectTerms Animal genetic engineering
Animals
Animals, Genetically Modified - physiology
Antibiotics
Biology and Life Sciences
Chromosomes
Design
Drosophila melanogaster - genetics
Engineering and Technology
Female
Females
Gene expression
Gene pool
Genetic aspects
Genetic crosses
Genetic engineering
Genetic Engineering - methods
Genetic research
Insect Control - methods
Insect sterilization
Insects
Male
Males
Medicine and Health Sciences
Methods
Models, Animal
Pest Control, Biological - methods
Pests
Phenotypes
Research and Analysis Methods
Sex chromosomes
Sex Chromosomes - genetics
Sex Determination Analysis - methods
Sex Determination Processes - genetics
Sex preselection
Sex ratio
Statistical analysis
Transgenes - genetics
X chromosomes
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Title Genetic engineering of sex chromosomes for batch cultivation of non-transgenic, sex-sorted males
URI https://www.ncbi.nlm.nih.gov/pubmed/33137115
https://www.proquest.com/docview/2479450573
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https://doaj.org/article/7f863e26bcf64dfa8e8deacbbdd53190
http://dx.doi.org/10.1371/journal.pgen.1009180
Volume 16
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