Efficient Ablation of Genes in Human Hematopoietic Stem and Effector Cells using CRISPR/Cas9.

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Efficient Ablation of Genes in Human Hematopoietic Stem and Effector Cells using CRISPR/Cas9. / Mandal, Pankaj K; Ferreira, Leonardo M R; Collins, Ryan; Meissner, Torsten B; Boutwell, Christian L; Friesen, Max; Vrbanac, Vladimir; Garrison, Brian S; Stortchevoi, Alexei; Bryder, David; Musunuru, Kiran; Brand, Harrison; Tager, Andrew M; Allen, Todd M; Talkowski, Michael E; Rossi, Derrick J; Cowan, Chad A.

In: Cell Stem Cell, Vol. 15, No. 5, 2014, p. 643-652.

Research output: Contribution to journalArticle

Harvard

Mandal, PK, Ferreira, LMR, Collins, R, Meissner, TB, Boutwell, CL, Friesen, M, Vrbanac, V, Garrison, BS, Stortchevoi, A, Bryder, D, Musunuru, K, Brand, H, Tager, AM, Allen, TM, Talkowski, ME, Rossi, DJ & Cowan, CA 2014, 'Efficient Ablation of Genes in Human Hematopoietic Stem and Effector Cells using CRISPR/Cas9.', Cell Stem Cell, vol. 15, no. 5, pp. 643-652. https://doi.org/10.1016/j.stem.2014.10.004

APA

Mandal, P. K., Ferreira, L. M. R., Collins, R., Meissner, T. B., Boutwell, C. L., Friesen, M., ... Cowan, C. A. (2014). Efficient Ablation of Genes in Human Hematopoietic Stem and Effector Cells using CRISPR/Cas9. Cell Stem Cell, 15(5), 643-652. https://doi.org/10.1016/j.stem.2014.10.004

CBE

Mandal PK, Ferreira LMR, Collins R, Meissner TB, Boutwell CL, Friesen M, Vrbanac V, Garrison BS, Stortchevoi A, Bryder D, Musunuru K, Brand H, Tager AM, Allen TM, Talkowski ME, Rossi DJ, Cowan CA. 2014. Efficient Ablation of Genes in Human Hematopoietic Stem and Effector Cells using CRISPR/Cas9. Cell Stem Cell. 15(5):643-652. https://doi.org/10.1016/j.stem.2014.10.004

MLA

Vancouver

Mandal PK, Ferreira LMR, Collins R, Meissner TB, Boutwell CL, Friesen M et al. Efficient Ablation of Genes in Human Hematopoietic Stem and Effector Cells using CRISPR/Cas9. Cell Stem Cell. 2014;15(5):643-652. https://doi.org/10.1016/j.stem.2014.10.004

Author

Mandal, Pankaj K ; Ferreira, Leonardo M R ; Collins, Ryan ; Meissner, Torsten B ; Boutwell, Christian L ; Friesen, Max ; Vrbanac, Vladimir ; Garrison, Brian S ; Stortchevoi, Alexei ; Bryder, David ; Musunuru, Kiran ; Brand, Harrison ; Tager, Andrew M ; Allen, Todd M ; Talkowski, Michael E ; Rossi, Derrick J ; Cowan, Chad A. / Efficient Ablation of Genes in Human Hematopoietic Stem and Effector Cells using CRISPR/Cas9. In: Cell Stem Cell. 2014 ; Vol. 15, No. 5. pp. 643-652.

RIS

TY - JOUR

T1 - Efficient Ablation of Genes in Human Hematopoietic Stem and Effector Cells using CRISPR/Cas9.

AU - Mandal, Pankaj K

AU - Ferreira, Leonardo M R

AU - Collins, Ryan

AU - Meissner, Torsten B

AU - Boutwell, Christian L

AU - Friesen, Max

AU - Vrbanac, Vladimir

AU - Garrison, Brian S

AU - Stortchevoi, Alexei

AU - Bryder, David

AU - Musunuru, Kiran

AU - Brand, Harrison

AU - Tager, Andrew M

AU - Allen, Todd M

AU - Talkowski, Michael E

AU - Rossi, Derrick J

AU - Cowan, Chad A

N1 - The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Stem Cell Aging (013212073)

PY - 2014

Y1 - 2014

N2 - Genome editing via CRISPR/Cas9 has rapidly become the tool of choice by virtue of its efficacy and ease of use. However, CRISPR/Cas9-mediated genome editing in clinically relevant human somatic cells remains untested. Here, we report CRISPR/Cas9 targeting of two clinically relevant genes, B2M and CCR5, in primary human CD4(+) T cells and CD34(+) hematopoietic stem and progenitor cells (HSPCs). Use of single RNA guides led to highly efficient mutagenesis in HSPCs but not in T cells. A dual guide approach improved gene deletion efficacy in both cell types. HSPCs that had undergone genome editing with CRISPR/Cas9 retained multilineage potential. We examined predicted on- and off-target mutations via target capture sequencing in HSPCs and observed low levels of off-target mutagenesis at only one site. These results demonstrate that CRISPR/Cas9 can efficiently ablate genes in HSPCs with minimal off-target mutagenesis, which could have broad applicability for hematopoietic cell-based therapy.

AB - Genome editing via CRISPR/Cas9 has rapidly become the tool of choice by virtue of its efficacy and ease of use. However, CRISPR/Cas9-mediated genome editing in clinically relevant human somatic cells remains untested. Here, we report CRISPR/Cas9 targeting of two clinically relevant genes, B2M and CCR5, in primary human CD4(+) T cells and CD34(+) hematopoietic stem and progenitor cells (HSPCs). Use of single RNA guides led to highly efficient mutagenesis in HSPCs but not in T cells. A dual guide approach improved gene deletion efficacy in both cell types. HSPCs that had undergone genome editing with CRISPR/Cas9 retained multilineage potential. We examined predicted on- and off-target mutations via target capture sequencing in HSPCs and observed low levels of off-target mutagenesis at only one site. These results demonstrate that CRISPR/Cas9 can efficiently ablate genes in HSPCs with minimal off-target mutagenesis, which could have broad applicability for hematopoietic cell-based therapy.

U2 - 10.1016/j.stem.2014.10.004

DO - 10.1016/j.stem.2014.10.004

M3 - Article

VL - 15

SP - 643

EP - 652

JO - Cell Stem Cell

T2 - Cell Stem Cell

JF - Cell Stem Cell

SN - 1934-5909

IS - 5

ER -