Supplementary Materials

Supplementary Material for:

CRISPR-Cas9 gene repair of hematopoietic stem cells from patients with X-linked chronic granulomatous disease

Suk See De Ravin,* Linhong Li, Xiaolin Wu, Uimook Choi, Cornell Allen, Sherry Koontz, Janet Lee, Narda Theobald-Whiting, Jessica Chu, Mary Garofalo, Colin Sweeney, Lela Kardava, Susan Moir, Angelia Viley, Pachai Natarajan, Ling Su, Douglas Kuhns, Kol A. Zarember, Madhusudan V. Peshwa, Harry L. Malech*

*Corresponding author. Email: sderavin{at}niaid.nih.gov (S.S.D.R.); hmalech{at}nih.gov (H.L.M.)

Published 11 January 2017, Sci. Transl. Med. 9, eaah3480 (2017)
DOI: 10.1126/scitranslmed.aah3480

This PDF file includes:

  • Fig. S1. Evaluation of the CRISPR/Cas9 system in an immortalized B cell line from a patient with the CYBB exon 7 C676T mutation.
  • Fig. S2. Effects of phosphorothioate modifications to a single-stranded oligonucleotide donor template for CRISPR-mediated gene repair in immortalized B cells derived from an X-CGD patient.
  • Fig. S3. Optimization of CRISPR/Cas9/single-guide RNA and single-stranded oligonucleotide delivery into human CD34+ HSPCs by electroporation.
  • Fig. S4. FACS analysis for gp91phox expression in CD3+ T cells.
  • Fig. S5. Sequencing of CYBB exon 7 in FACS-sorted human CD45+ cells derived from P1 CD34+ HSPCs after transplant into NSG mice.
  • Fig. S6. CRISPR-mediated correction of CD34+ HSPCs from a second X-CGD patient (P2).
  • Fig. S7. Peripheral blood from NSG mice 26 weeks after transplant with corrected P2 CD34+ HSPCs.
  • Fig. S8. Specificity of CRISPR/single-guide RNA for targeting the CYBB mutation.
  • Table S1. Deep sequencing of CYBB site in sorted human CD45+CD34+ cells from the bone marrow and spleen of NSG mice at 6.5 months after transplant of gene-corrected P2 CD34+ HSPCs.
  • Table S2. Potential alternate genomic target sites.
  • Table S3. Targeted sequencing of selected computationally predicted off-target sites.
  • Table S4. Evaluation of specificity of CRISPR/single-guide RNA by sequencing CD34+ HSPCs from healthy donors and two X-CGD patients using the same
    mutation-specific single-guide RNA2.

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