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Table 1 Summaries of genome editing studies performed on preclinical models of inborn errors of metabolism

From: Genome editing for inborn errors of metabolism: advancing towards the clinic

IEM Summary Comment Reference
Hemophilia B Targeting FIX disease locus
Dual AAV8 vectors:
Zinc-finger nucleases (ZFNs) Partial FIX cDNA
Neonatal, hemophilia B mouse model
First in vivo study using therapeutic genome editing
Low rates of homology-directed repair (HDR) detectable off-target events (<4%)
Li et al., 2011 [33]
Hemophilia B Targeting Alb safe harbor
Dual AAV8 vectors:
ZFNs
Partial FIX cDNA
Adult C57BL/6+ hemophilia B mouse models
Low rate of NHEJ-mediated correction (0.5% fused mRNA transcripts)
Due to integration in Alb, secretion of corrected protein led to therapeutic levels of circulating FIX
Duration of effect 12 weeks after single treatment
Sharma et al., 2015 [34]
Hemophilia B Targeting Alb safe harbor
No endonuclease
Single AAV8 vector targeting FIX donor as 2A fusion to Alb
Neonatal + adult hemophilia B mice
No off-target
Low rate of HDR (0.1% fused mRNA transcripts)
Addition of amino-terminal proline to FIX due to 2A peptide
Barzel et al., 2015 [35]
Hereditary Tyrosinemia Type I Targeting Fah disease locus
CRISPR: SpCas9/gRNA + ssDNA oligonucleotide
Naked DNA
Positive selection of hereditary tyrosinemia type I (HT-I) mouse model
Low HDR (0.4%); increased to 33.5% after 30 days
Off-target for Fah gRNA < 0.3% (NIH3T3 cells)
Yin et al., 2014 [81]
Hereditary Tyrosinemia Type I Targeting Fah disease locus
CRISPR: SpCas9 mRNA, LNP delivery (nano.Cas9)
FAH2 gRNA + donor (AAV-HDR)
Transient expression SpCas9 (LNP)
HDR 6% without selection
Low level off-target cutting (Hepa1-6 cells)
Yin et al., 2016 [37]
Hereditary Tyrosinemia Type I Targeting Hpd (disease locus)
CRISPR: SpCas9 + 2 gRNAs non-homologous end joining (NHEJ)
Naked DNA
Positive selection of HT-I mouse model
Metabolic reprogramming
Multiplex editing (2 guides)
8% NHEJ efficiency at both cut sites 1-week post
68% efficiency 4-weeks
post (+ selection)
Pankowicz et al., 2016 [39]
Hereditary Tyrosinemia Type I Targeting Alb safe harbor
No endonuclease
rAAV8: Hpd shRNA + hFIX
C57Bl/6 wild-type mice
Inducible positive selection using CEPHOBA
Initial homologous recombination < 1%; after selection 50%
Nygaard et al., 2016 [41]
Ornithine Transcarbamylase Deficiency Targeting Otc disease locus
CRISPR: SaCas9 + gRNA
Dual AAV8 vectors:
SaCas9 + gRNA-donor Otc mouse model
Smaller Cas9 orthologue
S. aureus
10% HDR in neonatal mice
Large deletions in adult mice ➔ hyperammonemia
Yang et al., 2016 [43]
Lysosomal Storage Disorders (MPSI, MPSII) Targeting Alb safe harbor
Dual AAV8 vectors:
ZFNs + donor
C57Bl/6 wild-type mice
Therapeutic protein detectable by Western blot
Phase I clinical trial MPSI:
3 AAV6 vectors:
ZFN + ZFN + donor
Sharma et al., 2015 [34]
Glycogen Storage Disorder Type Ia Targeting Rosa26 safe harbor
Dual AAV8/AAV9
ZFNs + donor
GSD1a mouse model
AAV8: Improvement in survival
AAV9: Improvement in biochemical phenotype
Positive selection of corrected hepatocytes
Landau et al., 2016 [44]