Broadening the GMO risk assessment in the EU for genome editing technologies in agriculture

Kawall, Katharina; Cotter, Janet; Then, Christoph

doi:10.1186/s12302-020-00361-2

Environmental Sciences Europe

Table 4 Examples of genome editing in farm animals for increased efficiency of production

From: Broadening the GMO risk assessment in the EU for genome editing technologies in agriculture

Animal	Target	Trait	Technological specification	References	Off-target analysis
Cow	Intraspecies POLLED allele substitution	No horns	TALENs (SDN-3)	[137]	WGS, no OTEs detected. But: additional integration of template plasmid discovered using whole genome sequencing data [76, 77]
Pig	Deletion of SRCR5 region of CD163	Resistance to PRRSV	CRISPR/Cas9 (SDN-1)	[229, 230] ^a	Altered cytokine levels (CCL3L1 and MIG)
Pig	Deletion of SRCR5 region of CD163	Resistance to PRRSV	CRISPR/Cas9 (SDN-1)	[231]	No analysis of OTEs
Pig	Knockins of antiviral small hairpin RNAs (shRNAs) at Rosa26 locus	Resistance to African swine fever	CRISPR/Cas9 (SDN-2)	[232]	Biased, screened 10 predicted off-target sites: no OTEs detected
Pig	Gene knockout of ANPEP (Amino peptidase N)	Resistance to TGEV	CRISPR/Cas9 (SDN-1)	[233]	No analysis of OTEs
Pig	Knockin of CRISPR/Cas9 targeting viral p30 gene	Resistance to TGEV	CRISPR/Cas9 (SDN-3)	[234]	No analysis of OTEs
Cow	Gene knockin of human lysozyme (hLYZ) gene to bovine β-casein locus	Increased antibacterial properties in their milk (reduced susceptibility to mastitis)	ZNF (SDN-3)	[235]	Biased, screened 10 predicted off-target sites: no OTEs detected But: random integration of transgenic fragments at an off-target site
Cow	Gene knockin of mouse SP110 gene	Increased resistance to tuberculosis	TALENs (SDN-3)	[236]	No analysis of OTEs
Cow	Gene knockin of bovine NRAMP-1 gene	Increased resistance to tuberculosis	CRISPR/Cas9 nickase (SDN-3)	[237]	Biased, screened 15 predicted off-target sites: no OTEs detected
Chicken	Gene knockout of ovomucoid (OVM)	Reduced allergenicity	CRISPR/Cas9 (SDN-1)	[238]	Biased, screened 3 predicted off-target sites: no OTEs detected. Analysis of plasmid integration: negative
Chicken	Gene knockout of ovalbumin (OV)	Reduced allergenicity	TALENs (SDN-1)	[239]	Biased, screened predicted off-target sites: no OTEs detected. Analysis of plasmid integration: negative

Target genes, traits and genome editing technique used to create genome-edited farm animals with increased efficiency of production are shown. The respective references were also screened for the analysis of off-target effects
CRISPR/Cas9, clustered regularly interspaced short palindromic repeats/CRISPR-associated 9; SDN-1, site-directed nuclease-1; TALENs, transcription activator-like effector nucleases; OTEs, off-target effects; ZNF, zinc finger nucleases
^aThese two studies relate to the development of the same genome-edited organism and are treated as one study

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