Table 1 Off-target effects in bone marrow cells following intravenous treatment of β-globin/GFP mice with γtcPNA4/donor DNA NPs.

From: In vivo correction of anaemia in β-thalassemic mice by γPNA-mediated gene editing with nanoparticle delivery

Gene locus

Sequences of partial homology (5′–3′)

Size of region sequenced

Alleles sequenced

Number modified

Frequency (%)

β-globin

TGCCCTGAAAGAAAGAGA

128

1,399,786

78,833

0.56

Vascular cell adhesion protein precursor 1

AGCCCTGAAAGAAAGAGA

111

480,013

0

0

Polypyrimidine tract binding protein

GAACCTGAAAGAAAGAGA

101

349,723

26

0.0074

Protocadherin fat 4 precursor

CACCCTGAAAGAAAGAAG

115

73,245

0

0

Olfactory receptor 266

AAGCCTGAAAGAAAGATT

172

1,092,990

2

0.00018

Syntaxin binding protein

AGAAATGAAAGAAAGAGA

150

2,478,636

0

0

Muscleblind like protein

GGTGGTGAAAGAAAGAGA

165

2,331,971

0

0

Ceruloplasmin isoform

AGGACTGAAAGAAAGAGT

154

1,390,439

0

0

Total off-target

  

8,197,017

28

0.00034

  1. The top seven gene loci in the mouse genome with partial homology to the 18 bp γtcPNA4 target site in β-globin intron 2 were identified, with the sequences as indicated. β-globin/GFP mice were treated with SCF followed by intravenous infusion with NPs containing γtcPNA4/donor DNA, and genomic DNA from c-Kit+ BM cells was subject to deep sequencing analysis at these loci. The size of the region sequenced around each site is listed, along with the number of alleles sequenced and the number of alleles with modified sequences.
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