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Efficient marmoset genome engineering by autologous embryo transfer and CRISPR/Cas9 technology. | LitMetric

Efficient marmoset genome engineering by autologous embryo transfer and CRISPR/Cas9 technology.

Sci Rep

Section of Animal Research and Laboratory of Animal Resources, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.

Published: October 2021

AI Article Synopsis

  • Genetic engineering of non-human primates like common marmosets is seen as a way to create ideal models for studying human genetic diseases.
  • A new method using autologous embryo transfer (AET) and CRISPR/Cas9 has been developed, allowing for faster production of genetically modified marmosets while adhering to humane experimental practices.
  • This technique successfully produced six marmosets with mutations in the FMR1 gene using only 18 female marmosets, demonstrating efficiency and reducing the number of animals needed for research.

Article Abstract

Genetic engineering of non-human primates, which are most closely related to humans, has been expected to generate ideal animal models for human genetic diseases. The common marmoset (Callithrix jacchus) is a non-human primate species adequate for the production of genetically modified animals because of their small body size and high reproductive capacity. Autologous embryo transfer (AET) is routinely utilized in assisted reproductive technologies for humans but not for experimental animals. This study has developed a novel method for efficiently producing mutant marmosets using AET and CRISPR/Cas9 systems. The embryos were recovered from oviducts of naturally mated females, injected with Cas9/guide RNA, and transferred into the oviducts of the donors. This AET method can reduce the time for in vitro culture of embryos to less than 30 min. This method uses an embryo donor as the recipient, thus reducing the number of animals and allowing for "Reduction" in the 3R principles of humane experimental technique. Furthermore, this method can utilize nulliparous females as well as parous females. We applied our novel method and generated the 6 marmosets carrying mutations in the fragile X mental retardation 1 (FMR1) gene using only 18 females including 14 nulliparous females.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8511084PMC
http://dx.doi.org/10.1038/s41598-021-99656-4DOI Listing

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