ArticleTransgenic research2017
Generation of gene edited birds in one generation using sperm transfection assisted gene editing (STAGE).
Article in Transgenic research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed, 64 citations in OpenAlex.
- Review
- Production of immune receptor knockout chickens via directAnimal biotechnology · 2025Article
- Genome Editing in the Chicken: From PGC-Mediated Germline Transmission to Advanced Applications.International journal of molecular sciences · 2025Review
- Advances in Genomics and Postgenomics in Poultry Science: Current Achievements and Future Directions.International journal of molecular sciences · 2025Review
- Application of CRISPR/Cas gene editing for infectious disease control in poultry.Open life sciences · 2025Review
- One-Step Genetic Modification by Embryonic Doral Aorta Injection of Adenoviral CRISPR/Cas9 Vector in Chicken.International journal of molecular sciences · 2024Article
- Identification of Two Potential Gene Insertion Sites for Gene Editing on the Chicken Z/W Chromosomes.Genes · 2024Article
- Hyperthermia suppresses the biological characteristics and migration of chicken primordial germ cells.Frontiers in genome editing · 2024Article
- Review
- Review
- Avian Sex Determination: A Chicken and Egg Conundrum.Sexual development : genetics, molecular biology, evolution, endocrinology, embryology, and pathology of sex determination and differentiation · 2023Review
- Genome editing of avian species: implications for animal use and welfare.Laboratory animals · 2022Article
- Genome editing approaches using reproductive cells/tissues in flowering plants.Frontiers in genome editing · 2022Review
- Genetic Regulation of Avian Testis Development.Genes · 2021Review
- Detection of a biolistic delivery of fluorescent markers and CRISPR/Cas9 to the pollen tube.Plant reproduction · 2021Article
- Potential of genomic technologies to improve disease resistance in molluscan aquaculture.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2021Review
- Advances in the Study of Olfaction in Eusocial Ants.Insects · 2021Review
- Primary sex determination in birds depends on DMRT1 dosage, but gonadal sex does not determine adult secondary sex characteristics.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Improving germline transmission efficiency in chimeric chickens using a multi-stage injection approach.PloS one · 2021Article
- Recent Advances in the Application of CRISPR/Cas9 Gene Editing System in Poultry Species.Frontiers in genetics · 2021Review
Corrections and comments
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Authors and funding
8 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Generating transgenic and gene edited mammals involves in vitro manipulation of oocytes or single cell embryos. Due to the comparative inaccessibility of avian oocytes and single cell embryos, novel protocols have been developed to produce transgenic and gene edited birds. While these protocols are relatively efficient, they involve two generation intervals before reaching complete somatic and germline expressing transgenic or gene edited birds. Most of this work has been done with chickens, and many protocols require in vitro culturing of primordial germ cells (PGCs). However, for many other bird species no methodology for long term culture of PGCs exists. Developing methodologies to produce germline transgenic or gene edited birds in the first generation would save significant amounts of time and resource. Furthermore, developing protocols that can be readily adapted to a wide variety of avian species would open up new research opportunities. Here we report a method using sperm as a delivery mechanism for gene editing vectors which we call sperm transfection assisted gene editing (STAGE). We have successfully used this method to generate GFP knockout embryos and chickens, as well as generate embryos with mutations in the doublesex and mab-3 related transcription factor 1 (DMRT1) gene using the CRISPR/Cas9 system. The efficiency of the method varies from as low as 0% to as high as 26% with multiple factors such as CRISPR guide efficiency and mRNA stability likely impacting the outcome. This straightforward methodology could simplify gene editing in many bird species including those for which no methodology currently exists.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.