Multiplex Genome Engineering Using CRISPR/Cas Systems

Details

• Alternative Title:
  Science
• Personal Author:
  Cong, Le ; Ran, F. Ann ; Cox, David ; Lin, Shuailiang ; Barretto, Robert ; Habib, Naomi ; Hsu, Patrick D. ; Wu, Xuebing ; Jiang, Wenyan ; Marraffini, Luciano A. ; Zhang, Feng
• Description:
  Functional elucidation of causal genetic variants and elements requires precise genome editing technologies. The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas adaptive immune system has been shown to facilitate RNA-guided site-specific DNA cleavage. We engineered two different type II CRISPR/Cas systems and demonstrate that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells. Cas9 can also be converted into a nicking enzyme to facilitate homology-directed repair with minimal mutagenic activity. Lastly, multiple guide sequences can be encoded into a single CRISPR array to enable simultaneous editing of several sites within the mammalian genome, demonstrating easy programmability and wide applicability of the RNA-guided nuclease technology.
• Subjects:
  Animals Article Base Sequence Caspase 9 DNA DNA Cleavage Genetic Engineering Genetic Loci Genome Humans Inverted Repeat Sequences Mice Microarray Analysis Molecular Sequence Data Mutagenesis Recombinational DNA Repair RNA Streptococcus Pyogenes

  More +
• Source:
  Science. 339(6121):819-823.
• Pubmed ID:
  23287718
• Pubmed Central ID:
  PMC3795411
• Document Type:
  Journal Article
• Funding:
  DP1 MH100706/MH/NIMH NIH HHS/United States ; DP1MH100706/DP/NCCDPHP CDC HHS/United States ; DP2 AI104556/AI/NIAID NIH HHS/United States ; DP2AI104556/AI/NIAID NIH HHS/United States ; R01 NS073124/NS/NINDS NIH HHS/United States ; R01-CA133404/CA/NCI NIH HHS/United States ; R01-GM34277/GM/NIGMS NIH HHS/United States ; T32 GM007753/GM/NIGMS NIH HHS/United States ; Howard Hughes Medical Institute/United States
• Volume:
  339
• Issue:
  6121
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:3c1fe915cd30007d56c534912eceb250d5716fb213a3ef53668f92ea176cad76
• Download URL:
  https://stacks.cdc.gov/view/cdc/29753/cdc_29753_DS1.pdf
• File Type:
  [PDF - 287.82 KB ]