Skip to main content
. 2021 Feb 1;10(3):513. doi: 10.3390/jcm10030513

Table 1.

HDR-mediated genome editing in HSPCs for inherited disorders.

Disease Study HSPC Source Gene Editing System Editing Efficiency at Target Locus Ref.
SCD In vitro BM, SCD subjects ZFN mRNA + IDLV 18.4% HBB [74]
BM, SCD subjects Cas9 mRNA + IDLV 20% HBB [75]
Blood, SCD subjects Cas9 RNP + ssODN 25% HBB [27]
mPB, SCD subjects Cas9 RNP + AAV6 50% HBB [79]
In vivo CB, healthy subjects ZFN mRNA + IDLV 0.21% HBB (1° NSG BM)
0.27% HBB (1° NSG spleen)		 [74]
mPB, healthy subjects Cas9 RNP + ssODN 2.3% HBB (1° NSG BM) [27]
mPB, healthy subjects Cas9 RNP + AAV6 90% HBB (1° NSG BM + selection)
3.5% HBB (1° NSG BM-selection)		 [79]
SCID-X1 In vitro CB, SCID-X1 subject ZFN mRNA + IDLV >20% IL2RG [80]
ZFN mRNA + AAV6 >50% IL2RG
mPB/CB, healthy subjects Cas9 RNP + AAV6 45% IL2RG [81]
mPB, SCID-X1 subjects Cas9 RNP + AAV6 44.5% IL2RG
In vivo CB, SCID-X1 subject ZFN mRNA + IDLV 5% IL2RG (1° NSG BM HSPCs) [80]
ZFN mRNA + AAV6 >25% IL2RG (1° NSG BM HSPCs)
CB, healthy subjects Cas9 RNP + AAV6 25.5% IL2RG (1° NSG BM)
9.5% to 20% IL2RG (2° NSG BM)		 [81]
mPB, SCID-X1 subjects Cas9 RNP + AAV6 Corrected IL2RG (1° NSG spleen)
X-CGD In vitro mPB, X-CGD subjects ZFN mRNA + AAV6 15% AAVS1, gp91ph°x expression in HSPC-derived myeloid cells [82]
mPB, X-CGD subjects Cas9 mRNA + ssODN 31% AAVS1, gp91ph°x expression in HSPC-derived myeloid cells [71]
In vivo mPB, X-CGD subjects ZFN mRNA + AAV6 10.7% AAVS1, gp91ph°x (1° NSG BM) [82]
mPB, X-CGD subjects Cas9 mRNA + ssODN 15.6% AAVS1, gp91ph°x (1° NSG PB)
16.5% AAVS1, gp91ph°x (1° NSG BM)		 [71]
XHIM In vitro mPB, XHIM subjects TALEN + AAV6 13.2% CD40LG, 5′ UTR [83]
Cas9 mRNA + AAV6 16.2% CD40LG, 5′ UTR
Cas9 RNP + AAV6 20.8% CD40LG, 5′ UTR
In vivo mPB, XHIM subjects TALEN + AAV6 ± adeno helper protein Average of all editing strategies:
4.4% CD40LG (1° NSG BM). No increased editing with addition of adeno helper protein		 [83]
Cas9 RNP + AAV6 ± adeno helper protein
SCN In vitro mPB, healthy subjects Cas9 RNP + AAV6 30% ELANE (exon 4 gRNA) [84]
BM, SCN subjects Cas9 RNP + AAV6 30% ELANE (exon 4 gRNA)
20% ELANE (L172P gRNA)
In vivo BM, SCN subjects Cas9 RNP + AAV6 3.1% ELANE-corrected neutrophils
(1° NOG-EXL BM)		 [84]
WAS In vitro mPB, healthy subjects Cas9 RNP + AAV6 69% WAS, bulk CD34 + cells
67.3% WAS, sorted HSCs (day 7)
58.8% WAS, sorted HSCs (day 14)		 [85]
mPB/BM, WAS subjects Cas9 RNP + AAV6 46.4% WAS, CD34 + cells (ddPCR)
45.5% WAS, CD34 + cells (flow)
In vivo mPB/BM, WAS subjects Cas9 RNP + AAV6 36.8% WAS (1° NSG BM)
16% WAS (2° NSG BM)		 [85]
LSD In vitro CB/mPB, healthy subjects Cas9 RNP + AAV6 28% CCR5 (PGK-IDUA) [86]
mPB/CB, healthy subjects Cas9 RNP + AAV6 ~50% HBA1 (LAL donor) [87]
In vivo mPB/CB, healthy subjects Cas9 RNP + AAV6 5–6% CCR5, PGK-IDUA (1° NSG BM) [86]
mPB/CB, healthy subjects Cas9 RNP + AAV6 ~8% HBA1 (1° NSG mice BM) [87]

Abbreviations: AAV6: adeno-associated virus 6; BM: bone marrow; PB: peripheral blood; IDLV: integration-deficient lentivirus; tNGFR: truncated nerve growth-factor receptor; ssODN: single-stranded oligonucleotide; RNP: ribonucleoprotein; IL2R: interleukin-2 receptor common gamma chain; CB:, cord blood; mPB: mobilized peripheral blood; UTR: untranslated region; NSG: NOD/SCID/IL-2rγnull; NOG: NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac; NOG-EXL: NOD.Cg-Prkdcscid Il2rgtm1Sug Tg(SV40/HTLV-IL3, CSF2)10–7Jic/JicTac; HBB: beta-globin gene; CYBB: cytochrome B-245 beta chain; CD40LG: CD40 ligand; CCR5: C-C chemokine receptor type 5; IDUA: alpha-L-iduronidase; LAL: lysosomal acid lipase; SCD: sickle cell disease; ZFN: zinc-finger nuclease; TALEN: transcription-activator like effector nuclease; SCID-X1: X-linked severe combined immuno-deficiency; X-CGD: X-linked chronic granulomatous disorder; XHIM: X-linked hyper immunoglobulin (Ig)M syndrome; SCN: severe congenital neutropenia; WAS: Wiskott–Aldrich syndrome; LSD: lysosomal storage disorders; HSPCs: hematopoietic stem/progenitor cells; HBA1: hemoglobin A1; PGK: phosphoglycerate kinase; ELANE: elastase, neutrophil expressed.