Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer...

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GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University of California, San Francisco, United States

Transcript of Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer...

Page 1: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

GENOME ENGINEERINGAlexendar Perez M.D., Ph.D

National Cancer Institute, NIH, United StatesDepartment of Anesthesia and Perioperative Care, University of California,

San Francisco, United States

Page 2: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Genome Engineering with Type II CRISPR Systems

Page 3: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Cas 9

Guide RNA

CRISPR Endonuclease

CRISPR Targeting Element

Page 4: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Cas 9

Guide RNA

CRISPR-Cas9 Targeting Complex

Page 5: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Cas 9

Guide RNA

Chromosome

Target

Page 6: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Cas 9

Complement

Chromosome

Target PAM

tracrRNA

tracrRNA + Complement = Guide RNA

Page 7: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Cas 9

Complement

Chromosome

Target PAM

tracrRNA

tracrRNA + Complement = Guide RNA

Page 8: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Chromosome Part A Chromosome Part B

CRISPR Engineered DNA Double Strand Break

Page 9: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Genome Engineering Occurs Through Repair of DNA Double Strand Break

Page 10: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Chromosome Part A Chromosome Part B

CRISPR Precise Modification of Genomic Locus

Modifying DNA

Page 11: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Chromosome Part A Chromosome Part B

Chromosome

Modifying DNA

Precision Modified Genomic Locus

Page 12: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Chromosome Part A Chromosome Part B

CRISPR Disrupted Modification of Genomic Locus

Blunted DNA

Page 13: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Chromosome Part A Chromosome Part B

Blunted DNA

Chromosome Part A Chromosome Part B

Page 14: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Chromosome Part A Chromosome Part B

Blunted DNA

Chromosome Part A Chromosome Part B

Chromosome

Disruption Modified Genomic Locus

Page 15: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Chromosome

Target

Unique Guide RNA Target

Page 16: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Chromosome

Target Target Target Target

Unique Guide RNA Target?

Page 17: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Genomic Evisceration

Page 18: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Perez et al. 2017 Nature Biotechnology

Page 19: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Chromosome

Multi-Chromosome Targeting Guide RNA

Chromosome

Target

Target

Page 20: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Chromosome

Chromosomal Translocations

Chromosome

Page 21: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Chromosome

Chromosomal TranslocationsHallmark of Cancer

Chromosome

Page 22: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Perez et al. 2017 Nature Biotechnology

Page 23: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

GuideScan Software Guarantees Guide RNA Uniqueness and Specificity

Page 24: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

Perez et al. 2017 Nature Biotechnology

Page 25: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

GuideScan ensures gRNA Specificity

Page 26: Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer Institute, NIH, United States Department of Anesthesia and Perioperative Care, University

ACKNOWLEDGEMENTSVidigal Laboratory NCI/NIH• Joana Vidigal, Ph.D• Mahendra Prajapat Ph.D.• Laura Sala Ph.D• Srividya Chandrasekhar

UCSF Department of Anesthesia and Perioperative Care• Kristina Sullivan, M.D.• Manny Pardo, M.D.• Judith Hellman, M.D.• Michael Gropper M.D., Ph.D

GITEX Technology Week• Welma Williams• United Arab Emirates

Memorial Sloan Kettering Cancer Center • Christina Leslie, Ph.D and Laboratory• Andrea Ventura, M.D., Ph.D and Laboratory


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