Table 3 Deep sequencing analysis of targeted gene editing versus off-target effects in human CD34+ haematopoietic cells following ex vivo treatment with SCF and 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

12,489,910

606,230

5.02

Serine Theronine Kinase

ATTCCTGAAAGAAAGCAC

189

3,330,879

0

0

Anoctamin-3

AATTCTGAAAGAAAGACC

150

5,805,518

1

0.000017

39s Ribosomal protein L17

AGCCCTGAAAGAATACCA

169

4,211,251

0

0

Neuroblast differentiation associated

TCCCTGAAAGAAAAAAGA

198

3,579,389

2

0.000055

Transcription enhance factor TEF1

TCTCCCTGAAAGAAAAAA

244

80,548

0

0

Rho GTPase activating protein

CAACATGAAAGAAAGAGA

154

8,256,220

0

0

Total off-target

  

25,263,805

3

0.000012

  1. The top six gene loci in the human genome with partial homology to the 18 bp γtcPNA4 target site in β-globin intron 2 were identified, with the sequences as indicated. Human CD34+ haematopoietic cells were treated ex vivo with SCF and with NPs containing γtcPNA4/donor DNA, and 2 days later genomic DNA from the 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.
Back to article page