Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer...
Transcript of Perez Alexendar GITEX 2019 Brief · GENOME ENGINEERING Alexendar Perez M.D., Ph.D National Cancer...
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
Genome Engineering with Type II CRISPR Systems
Cas 9
Guide RNA
CRISPR Endonuclease
CRISPR Targeting Element
Cas 9
Guide RNA
CRISPR-Cas9 Targeting Complex
Cas 9
Guide RNA
Chromosome
Target
Cas 9
Complement
Chromosome
Target PAM
tracrRNA
tracrRNA + Complement = Guide RNA
Cas 9
Complement
Chromosome
Target PAM
tracrRNA
tracrRNA + Complement = Guide RNA
Chromosome Part A Chromosome Part B
CRISPR Engineered DNA Double Strand Break
Genome Engineering Occurs Through Repair of DNA Double Strand Break
Chromosome Part A Chromosome Part B
CRISPR Precise Modification of Genomic Locus
Modifying DNA
Chromosome Part A Chromosome Part B
Chromosome
Modifying DNA
Precision Modified Genomic Locus
Chromosome Part A Chromosome Part B
CRISPR Disrupted Modification of Genomic Locus
Blunted DNA
Chromosome Part A Chromosome Part B
Blunted DNA
Chromosome Part A Chromosome Part B
Chromosome Part A Chromosome Part B
Blunted DNA
Chromosome Part A Chromosome Part B
Chromosome
Disruption Modified Genomic Locus
Chromosome
Target
Unique Guide RNA Target
Chromosome
Target Target Target Target
Unique Guide RNA Target?
Genomic Evisceration
Perez et al. 2017 Nature Biotechnology
Chromosome
Multi-Chromosome Targeting Guide RNA
Chromosome
Target
Target
Chromosome
Chromosomal Translocations
Chromosome
Chromosome
Chromosomal TranslocationsHallmark of Cancer
Chromosome
Perez et al. 2017 Nature Biotechnology
GuideScan Software Guarantees Guide RNA Uniqueness and Specificity
Perez et al. 2017 Nature Biotechnology
GuideScan ensures gRNA Specificity
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