A CRISPR-Cas9 delivery system for in vivo screening of genes in the immune system
Therapies that target the function of immune cells have significant clinical efficacy in diseases such as cancer and autoimmunity. Although functional genomics has accelerated therapeutic target discovery in cancer, its use in primary immune cells is limited because vector delivery is inefficient an...
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| Veröffentlicht in: | Nature communications Jg. 10; H. 1; S. 1668 - 10 |
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| Sprache: | Englisch |
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10.04.2019
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| Abstract | Therapies that target the function of immune cells have significant clinical efficacy in diseases such as cancer and autoimmunity. Although functional genomics has accelerated therapeutic target discovery in cancer, its use in primary immune cells is limited because vector delivery is inefficient and can perturb cell states. Here we describe CHIME: CHimeric IMmune Editing, a CRISPR-Cas9 bone marrow delivery system to rapidly evaluate gene function in innate and adaptive immune cells in vivo without ex vivo manipulation of these mature lineages. This approach enables efficient deletion of genes of interest in major immune lineages without altering their development or function. We use this approach to perform an in vivo pooled genetic screen and identify Ptpn2 as a negative regulator of CD8
+
T cell-mediated responses to LCMV Clone 13 viral infection. These findings indicate that this genetic platform can enable rapid target discovery through pooled screening in immune cells in vivo.
The use of functional genomics in primary immune cells has been limited by inefficient vector delivery and risk of perturbing cell states. Here the authors present CHimeric IMmune Editing (CHIME) for in vivo evaluation of gene function and pooled screening approaches. |
|---|---|
| AbstractList | Therapies that target the function of immune cells have significant clinical efficacy in diseases such as cancer and autoimmunity. Although functional genomics has accelerated therapeutic target discovery in cancer, its use in primary immune cells is limited because vector delivery is inefficient and can perturb cell states. Here we describe CHIME: CHimeric IMmune Editing, a CRISPR-Cas9 bone marrow delivery system to rapidly evaluate gene function in innate and adaptive immune cells in vivo without ex vivo manipulation of these mature lineages. This approach enables efficient deletion of genes of interest in major immune lineages without altering their development or function. We use this approach to perform an in vivo pooled genetic screen and identify Ptpn2 as a negative regulator of CD8
+
T cell-mediated responses to LCMV Clone 13 viral infection. These findings indicate that this genetic platform can enable rapid target discovery through pooled screening in immune cells in vivo.
The use of functional genomics in primary immune cells has been limited by inefficient vector delivery and risk of perturbing cell states. Here the authors present CHimeric IMmune Editing (CHIME) for in vivo evaluation of gene function and pooled screening approaches. The use of functional genomics in primary immune cells has been limited by inefficient vector delivery and risk of perturbing cell states. Here the authors present CHimeric IMmune Editing (CHIME) for in vivo evaluation of gene function and pooled screening approaches. Therapies that target the function of immune cells have significant clinical efficacy in diseases such as cancer and autoimmunity. Although functional genomics has accelerated therapeutic target discovery in cancer, its use in primary immune cells is limited because vector delivery is inefficient and can perturb cell states. Here we describe CHIME: CHimeric IMmune Editing, a CRISPR-Cas9 bone marrow delivery system to rapidly evaluate gene function in innate and adaptive immune cells in vivo without ex vivo manipulation of these mature lineages. This approach enables efficient deletion of genes of interest in major immune lineages without altering their development or function. We use this approach to perform an in vivo pooled genetic screen and identify Ptpn2 as a negative regulator of CD8+ T cell-mediated responses to LCMV Clone 13 viral infection. These findings indicate that this genetic platform can enable rapid target discovery through pooled screening in immune cells in vivo.Therapies that target the function of immune cells have significant clinical efficacy in diseases such as cancer and autoimmunity. Although functional genomics has accelerated therapeutic target discovery in cancer, its use in primary immune cells is limited because vector delivery is inefficient and can perturb cell states. Here we describe CHIME: CHimeric IMmune Editing, a CRISPR-Cas9 bone marrow delivery system to rapidly evaluate gene function in innate and adaptive immune cells in vivo without ex vivo manipulation of these mature lineages. This approach enables efficient deletion of genes of interest in major immune lineages without altering their development or function. We use this approach to perform an in vivo pooled genetic screen and identify Ptpn2 as a negative regulator of CD8+ T cell-mediated responses to LCMV Clone 13 viral infection. These findings indicate that this genetic platform can enable rapid target discovery through pooled screening in immune cells in vivo. Therapies that target the function of immune cells have significant clinical efficacy in diseases such as cancer and autoimmunity. Although functional genomics has accelerated therapeutic target discovery in cancer, its use in primary immune cells is limited because vector delivery is inefficient and can perturb cell states. Here we describe CHIME: CHimeric IMmune Editing, a CRISPR-Cas9 bone marrow delivery system to rapidly evaluate gene function in innate and adaptive immune cells in vivo without ex vivo manipulation of these mature lineages. This approach enables efficient deletion of genes of interest in major immune lineages without altering their development or function. We use this approach to perform an in vivo pooled genetic screen and identify Ptpn2 as a negative regulator of CD8 + T cell-mediated responses to LCMV Clone 13 viral infection. These findings indicate that this genetic platform can enable rapid target discovery through pooled screening in immune cells in vivo. Therapies that target the function of immune cells have significant clinical efficacy in diseases such as cancer and autoimmunity. Although functional genomics has accelerated therapeutic target discovery in cancer, its use in primary immune cells is limited because vector delivery is inefficient and can perturb cell states. Here we describe CHIME: CHimeric IMmune Editing, a CRISPR-Cas9 bone marrow delivery system to rapidly evaluate gene function in innate and adaptive immune cells in vivo without ex vivo manipulation of these mature lineages. This approach enables efficient deletion of genes of interest in major immune lineages without altering their development or function. We use this approach to perform an in vivo pooled genetic screen and identify Ptpn2 as a negative regulator of CD8+ T cell-mediated responses to LCMV Clone 13 viral infection. These findings indicate that this genetic platform can enable rapid target discovery through pooled screening in immune cells in vivo. The use of functional genomics in primary immune cells has been limited by inefficient vector delivery and risk of perturbing cell states. Here the authors present CHimeric IMmune Editing (CHIME) for in vivo evaluation of gene function and pooled screening approaches. Therapies that target the function of immune cells have significant clinical efficacy in diseases such as cancer and autoimmunity. Although functional genomics has accelerated therapeutic target discovery in cancer, its use in primary immune cells is limited because vector delivery is inefficient and can perturb cell states. Here we describe CHIME: CHimeric IMmune Editing, a CRISPR-Cas9 bone marrow delivery system to rapidly evaluate gene function in innate and adaptive immune cells in vivo without ex vivo manipulation of these mature lineages. This approach enables efficient deletion of genes of interest in major immune lineages without altering their development or function. We use this approach to perform an in vivo pooled genetic screen and identify Ptpn2 as a negative regulator of CD8 T cell-mediated responses to LCMV Clone 13 viral infection. These findings indicate that this genetic platform can enable rapid target discovery through pooled screening in immune cells in vivo. |
| ArticleNumber | 1668 |
| Author | Coxe, Matthew A. Weiss, Sarah A. Doench, John G. Nguyen, Thao H. Yates, Kathleen B. LaFleur, Martin W. Gillis, Jacob E. Coxe, Daniel J. Haining, W. Nicholas Sharpe, Arlene H. Trombley, Justin D. Brown, Flavian D. |
| Author_xml | – sequence: 1 givenname: Martin W. orcidid: 0000-0002-5017-774X surname: LaFleur fullname: LaFleur, Martin W. organization: Department of Microbiology and Immunobiology, Harvard Medical School, Department of Pediatric Oncology, Dana-Farber Cancer Institute, Evergrande Center for Immunological Diseases, Harvard Medical School and Brigham and Women’s Hospital – sequence: 2 givenname: Thao H. surname: Nguyen fullname: Nguyen, Thao H. organization: Department of Microbiology and Immunobiology, Harvard Medical School, Evergrande Center for Immunological Diseases, Harvard Medical School and Brigham and Women’s Hospital – sequence: 3 givenname: Matthew A. surname: Coxe fullname: Coxe, Matthew A. organization: Department of Microbiology and Immunobiology, Harvard Medical School, Evergrande Center for Immunological Diseases, Harvard Medical School and Brigham and Women’s Hospital – sequence: 4 givenname: Kathleen B. orcidid: 0000-0002-7383-5573 surname: Yates fullname: Yates, Kathleen B. organization: Department of Pediatric Oncology, Dana-Farber Cancer Institute, Broad Institute of Harvard and Massachusetts Institute of Technology – sequence: 5 givenname: Justin D. orcidid: 0000-0002-3856-3138 surname: Trombley fullname: Trombley, Justin D. organization: Department of Microbiology and Immunobiology, Harvard Medical School, Evergrande Center for Immunological Diseases, Harvard Medical School and Brigham and Women’s Hospital – sequence: 6 givenname: Sarah A. orcidid: 0000-0002-4155-0346 surname: Weiss fullname: Weiss, Sarah A. organization: Department of Pediatric Oncology, Dana-Farber Cancer Institute – sequence: 7 givenname: Flavian D. surname: Brown fullname: Brown, Flavian D. organization: Department of Microbiology and Immunobiology, Harvard Medical School, Department of Pediatric Oncology, Dana-Farber Cancer Institute, Evergrande Center for Immunological Diseases, Harvard Medical School and Brigham and Women’s Hospital – sequence: 8 givenname: Jacob E. surname: Gillis fullname: Gillis, Jacob E. organization: Department of Microbiology and Immunobiology, Harvard Medical School, Evergrande Center for Immunological Diseases, Harvard Medical School and Brigham and Women’s Hospital – sequence: 9 givenname: Daniel J. surname: Coxe fullname: Coxe, Daniel J. organization: School of Energy, Matter, and Transport Engineering at Arizona State University – sequence: 10 givenname: John G. orcidid: 0000-0002-3707-9889 surname: Doench fullname: Doench, John G. organization: Broad Institute of Harvard and Massachusetts Institute of Technology – sequence: 11 givenname: W. Nicholas orcidid: 0000-0001-7871-3762 surname: Haining fullname: Haining, W. Nicholas email: Nicholas_Haining@dfci.harvard.edu organization: Department of Pediatric Oncology, Dana-Farber Cancer Institute, Broad Institute of Harvard and Massachusetts Institute of Technology – sequence: 12 givenname: Arlene H. surname: Sharpe fullname: Sharpe, Arlene H. email: Arlene_Sharpe@hms.harvard.edu organization: Department of Microbiology and Immunobiology, Harvard Medical School, Evergrande Center for Immunological Diseases, Harvard Medical School and Brigham and Women’s Hospital, Broad Institute of Harvard and Massachusetts Institute of Technology |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/30971695$$D View this record in MEDLINE/PubMed |
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| PublicationYear | 2019 |
| Publisher | Nature Publishing Group UK Nature Publishing Group Nature Portfolio |
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| Snippet | Therapies that target the function of immune cells have significant clinical efficacy in diseases such as cancer and autoimmunity. Although functional genomics... The use of functional genomics in primary immune cells has been limited by inefficient vector delivery and risk of perturbing cell states. Here the authors... |
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