Mammalian interspecies substitution of immune modulatory alleles by genome editing.

Simon G Lillico,Lei Zhang,Edward J Rebar,David E Paschon,Alan J Mileham,Wenfang Tan,Rachel Mardjuki,C Bruce A Whitelaw,Fyodor D Urnov,Chris Proudfoot,David G Mclaren,Tim J King

doi:10.1038/srep21645

Abstract

We describe a fundamentally novel feat of animal genetic engineering: the precise and efficient substitution of an agronomic haplotype into a domesticated species. Zinc finger nuclease in-embryo editing of the RELA locus generated live born domestic pigs with the warthog RELA orthologue, associated with resilience to African Swine Fever. The ability to efficiently achieve interspecies allele introgression in one generation opens unprecedented opportunities for agriculture and basic research.

Highlights

We describe a fundamentally novel feat of animal genetic engineering: the precise and efficient substitution of an agronomic haplotype into a domesticated species
The domestic pig is highly susceptible to infection by African Swine Fever Virus, in contrast to present-day pig species found in Africa
The editing efficiency driven by the RELA-directed zinc finger nuclease (ZFN) nearly doubled what was seen with ZFNs successfully used to obtain live pigs bearing a disruption of the GGTA gene[11], suggesting that these nucleases may be well-suited for in-embryo editing

Summary

Introduction

We describe a fundamentally novel feat of animal genetic engineering: the precise and efficient substitution of an agronomic haplotype into a domesticated species. The development of targeted genome editing[1], pioneered by the zinc finger nuclease (ZFN) approach[2], enables variation not present within a given population to be used. This approach relies on engineered nucleases to direct a double-strand break (DSB) to a specific target sequence. When used in combination with an investigator-provided DNA template, specified changes can be introduced into the chromosome in a homology-based process during repair of the DSB In this way the original target sequence can be exchanged for a new sequence[3], enabling single allele introgression into the target animal population in one generation[4]. We observe single-step biallelic haplotype transfer by genotyping of piglets

Methods

Results

Conclusion