Gene-edited pigs are protected from porcine reproductive and respiratory syndrome virus

Whitworth, Kristin M.; Rowland, Raymond R. R.; Ewen, Catherine; Trible, Benjamin R.; Kerrigan, Maureen; Cino‐Ozuna, Ada G.; Samuel, Melissa; Lightner, Jonathan; McLaren, David G.; Mileham, Alan J.; Wells, Kevin D.; Prather, Randall S.

doi:10.1038/nbt.3434

Cited by 407 publications

(336 citation statements)

References 16 publications

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“…In previous work, we demonstrated that GM pigs that lack CD163 expression on macrophages fail to support the replication of a single type 2 PRRSV isolate (16). In the present study, we demonstrated that macrophages from KO pigs were resistant to infection with a large panel of type 1 and type 2 isolates ( Table 2), suggesting that all PRRSV isolates, regardless of genotype or clade, require CD163.…”

Section: Discussionmentioning

confidence: 52%

“…Recently, we showed that GM pigs lacking CD163 expression on macrophages failed to support infection with a type 2 virus (16). The purpose of the current study was to determine if CD163 knockout (KO) pigs were resistant to type 1 PRRSV genotype viruses and, second, to determine if the replacement of SRCR 5 with an hCD163L1 peptide sequence would be sufficient to confer on pigs resistance to both type 1 and type 2 viruses.…”

mentioning

confidence: 99%

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Replacement of Porcine CD163 Scavenger Receptor Cysteine-Rich Domain 5 with a CD163-Like Homolog Confers Resistance of Pigs to Genotype 1 but Not Genotype 2 Porcine Reproductive and Respiratory Syndrome Virus

Wells

Bardot

Whitworth

et al. 2017

J Virol

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CD163 knockout (KO) pigs are resistant to infection with genotype 2 (type 2) porcine reproductive and respiratory syndrome virus (PRRSV). Furthermore, the substitution of CD163 scavenger receptor cysteine-rich (SRCR) domain 5 with a homolog of human CD163-like (hCD163L1) SRCR 8 domain confers resistance of transfected HEK cells to type 1 PRRSV. As a means to understand the role of domain 5 in PRRSV infection with both type 1 and type 2 viruses, pigs were genetically modified (GM) to possess one of the following genotypes: complete knockout (KO) of CD163, deletions within SRCR domain 5, or replacement (domain swap) of SRCR domain 5 with a synthesized exon encoding a homolog of hCD163L1 SRCR domain 8. Immunophenotyping of porcine alveolar macrophages (PAMs) showed that pigs with the KO or SRCR domain 5 deletion did not express CD163. When placed in culture, PAMs from pigs with the CD163 KO phenotype were completely resistant to a panel consisting of six type 1 and nine type 2 isolates. PAMs from pigs that possessed the hCD163L1 domain 8 homolog expressed CD163 and supported the replication of all type 2 isolates, but no type 1 viruses. Infection of CD163-modified pigs with representative type 1 and type 2 viruses confirmed the in vitro results. The results confirm that CD163 is the likely receptor for all PRRS viruses. Even though type 1 and type 2 viruses are considered phenotypically similar at several levels, there is a distinct difference between the viral genotypes in the recognition of CD163.IMPORTANCE Genetic modification of the CD163 gene creates the opportunity to develop production animals that are resistant to PRRS, the costliest viral disease to ever face the swine industry. The results create further opportunities to develop refinements in the modification of CD163 with the goal of making pigs refractory to infection while retaining important CD163 functions.

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Section: Discussionmentioning

confidence: 52%

mentioning

confidence: 99%

Replacement of Porcine CD163 Scavenger Receptor Cysteine-Rich Domain 5 with a CD163-Like Homolog Confers Resistance of Pigs to Genotype 1 but Not Genotype 2 Porcine Reproductive and Respiratory Syndrome Virus

Wells

Bardot

Whitworth

et al. 2017

J Virol

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“…However, one study showed that Sn gene knockout pigs were infected by PRRSV as usual (49), and a recent study using geneedited pigs demonstrates the indispensability of pCD163 (28). Therefore, pCD163 is a core receptor for PRRSV infection and requires more in-depth research.…”

Section: Discussionmentioning

confidence: 99%

The Crystal Structure of the Fifth Scavenger Receptor Cysteine-Rich Domain of Porcine CD163 Reveals an Important Residue Involved in Porcine Reproductive and Respiratory Syndrome Virus Infection

Jiang

Qiao

et al. 2017

J Virol

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Porcine reproductive and respiratory syndrome (PRRS) has become an economically critical factor in swine industry since its worldwide spread in the 1990s. Infection by its causative agent, PRRS virus (PRRSV), was proven to be mediated by an indispensable receptor, porcine CD163 (pCD163), and the fifth scavenger receptor cysteine-rich domain (SRCR5) is essential for virus infection. However, the structural details and specific residues of pCD163 SRCR5 involved in infection have not been defined yet. In this study, we prepared recombinant pCD163 SRCR5 in Drosophila melanogaster Schneider 2 (S2) cells and determined its crystal structure at a high resolution of 2.0 Å. This structure includes a markedly long loop region and shows a special electrostatic potential, and these are significantly different from those of other members of the scavenger receptor cysteine-rich superfamily (SRCR-SF). Subsequently, we carried out structure-based mutational studies to identify that the arginine residue at position 561 (Arg561) in the long loop region is important for PRRSV infection. Further, we showed Arg561 probably takes effect on the binding of pCD163 to PRRSV during virus invasion. Altogether the current work provides the first view of the CD163 SRCR domain, expands our knowledge of the invasion mechanism of PRRSV, and supports a molecular basis for prevention and control of the virus.IMPORTANCE PRRS has caused huge economic losses to pig farming. The syndrome is caused by PRRSV, and PRRSV infection has been shown to be mediated by host cell surface receptors. One of them, pCD163, is especially indispensable, and its SRCR5 domain has been further demonstrated to play a significant role in virus infection. However, its structural details and the residues involved in infection are unknown. In this study, we determined the crystal structure of pCD163 SRCR5 and then carried out site-directed mutational studies based on the crystal structure to elucidate which residue is important. Our work not only provides structural information on the CD163 SRCR domain for the first time but also indicates the molecular mechanism of PRRSV infection and lays a foundation for future applications in prevention and control of PRRS.KEYWORDS CD163, Drosophila S2 cells, PRRSV, SRCR5, crystal structure P orcine reproductive and respiratory syndrome (PRRS) was first reported in 1987 in the United States (1) and has become a critical economic factor in the swine industry since its worldwide spread in the 1990s (2, 3). PRRS is characterized by

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“…A promising first use of gene editing in the dairy industry to address welfare issues may be the production of hornless dairy cattle with the introduction of the POLLED allele, which is nearly fixed in some beef breeds but low in frequency in Holsteins (Carlson et al, 2016). Other animal agriculture species are also making headway in the production of edited species, including pigs resistant to porcine reproductive and respiratory syndrome virus that are now being produced (Whitworth et al, 2016). Advances in the understanding of the influence of various genes and genomic variants on phenotypes will bring forth many future opportunities for the improvement of quantitative traits through gene editing.…”

Section: Genome Editingmentioning

confidence: 99%

Invited review: Reproductive and genomic technologies to optimize breeding strategies for genetic progress in dairy cattle

Fleming¹,

Abdalla²,

Maltecca

et al. 2018

Arch. Anim. Breed.

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Abstract. Dairy cattle breeders have exploited technological advances that have emerged in the past in regards to reproduction and genomics. The implementation of such technologies in routine breeding programs has permitted genetic gains in traditional milk production traits as well as, more recently, in low-heritability traits like health and fertility. As demand for dairy products increases, it is important for dairy breeders to optimize the use of available technologies and to consider the many emerging technologies that are currently being investigated in various fields. Here we review a number of technologies that have helped shape dairy breeding programs in the past and present, along with those potentially forthcoming. These tools have materialized in the areas of reproduction, genotyping and sequencing, genetic modification, and epigenetics. Although many of these technologies bring encouraging opportunities for genetic improvement of dairy cattle populations, their applications and benefits need to be weighed with their impacts on economics, genetic diversity, and society.

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Gene-edited pigs are protected from porcine reproductive and respiratory syndrome virus

Cited by 407 publications

References 16 publications

Replacement of Porcine CD163 Scavenger Receptor Cysteine-Rich Domain 5 with a CD163-Like Homolog Confers Resistance of Pigs to Genotype 1 but Not Genotype 2 Porcine Reproductive and Respiratory Syndrome Virus

Replacement of Porcine CD163 Scavenger Receptor Cysteine-Rich Domain 5 with a CD163-Like Homolog Confers Resistance of Pigs to Genotype 1 but Not Genotype 2 Porcine Reproductive and Respiratory Syndrome Virus

The Crystal Structure of the Fifth Scavenger Receptor Cysteine-Rich Domain of Porcine CD163 Reveals an Important Residue Involved in Porcine Reproductive and Respiratory Syndrome Virus Infection

Invited review: Reproductive and genomic technologies to optimize breeding strategies for genetic progress in dairy cattle

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