Successful cloning of coyotes through interspecies somatic cell nuclear transfer using domestic dog oocytes

Abstract

Interspecies somatic cell nuclear transfer (iSCNT) is an emerging assisted reproductive technology (ART) for preserving Nature's diversity. The scarcity of oocytes from some species makes utilisation of readily available oocytes inevitable. In the present study, we describe the successful cloning of coyotes (Canis latrans) through iSCNT using oocytes from domestic dogs (Canis lupus familiaris or dingo). Transfer of 320 interspecies-reconstructed embryos into 22 domestic dog recipients resulted in six pregnancies, from which eight viable offspring were delivered. Fusion rate and cloning efficiency during iSCNT cloning of coyotes were not significantly different from those observed during intraspecies cloning of domestic dogs. Using neonatal fibroblasts as donor cells significantly improved the cloning efficiency compared with cloning using adult fibroblast donor cells (P<0.05). The use of domestic dog oocytes in the cloning of coyotes in the present study holds promise for cloning other endangered species in the Canidae family using similar techniques. However, there are still limitations of the iSCNT technology, as demonstrated by births of morphologically abnormal coyotes and the clones' inheritance of maternal domestic dog mitochondrial DNA.

First cloned Bactrian camel (Camelus bactrianus) calf produced by interspecies somatic cell nuclear transfer: A step towards preserving the critically endangered wild Bactrian camels

“…Similarly, coyote pups ( Canis latrans ) were produced using dog ( Canis lupus familiaris ) oocytes [17] and a gaur calf ( Bos gaurus ) was produced from embryos reconstructed with bovine ( Bos taurus ) oocytes [8]. Even iSCNT embryos reconstructed from mouflon ( Ovis orientalis musimon ) donor cells and sheep ( Ovis aries ) oocytes [10] developed to term.…”

The potential and pitfalls of de‐extinction

“…This technology has triumphed only when using very closely related animals, with other cases including cloning of the coyote ( Canis latrans ) using domestic dog ( Canis lupus familiaris ) cells and surrogates (Hwang et al . ), wild cat ( Felis silvestris lybica ) and sand cat ( Felis margarita ) using the domestic cat ( Felis catus ; Gómez et al . , ), and a mouflon ( Ovis orientalis musimon ), an endangered subspecies of sheep, using domestic sheep ( Ovis aries ) oocyte and surrogate following nucleus retrieval from a cell of a deceased mouflon (Loi et al .…”

“…Cloning by iSCNT involves the transfer of a somatic cell nucleus to the enucleated oocyte of a different species or subspecies, and would be the method used for de-extinction due to the obvious impossibility of using the same species as a surrogate. This technology has triumphed only when using very closely related animals, with other cases including cloning of the coyote (Canis latrans) using domestic dog (Canis lupus familiaris) cells and surrogates (Hwang et al 2013), wild cat (Felis silvestris lybica) and sand cat (Felis margarita) using the domestic cat (Felis catus; G omez et al 2004,2008), and a mouflon (Ovis orientalis musimon), an endangered subspecies of sheep, using domestic sheep (Ovis aries) oocyte and surrogate following nucleus retrieval from a cell of a deceased mouflon (Loi et al 2001). It is clear, therefore, that deextinction by cloning using this technique is almost certainly achievable in theory.…”

Cross‐species cloning: influence of cytoplasmic factors on development

“…) and the coyote (Hwang et al . ), by using the oocytes from closely related species. It is commonly reported that the cloned animals are identical to their nuclear donors in genotypes and phenotypes, indicating the significant dominance of the nuclear genome in phenotypic determination.…”