RNA-directed Cleavage and Modification of DNA using CasX (CRISPR-CasX)

Tech ID: 26042 / UC Case 2017-016-0

Patent Status

Country Type Number Dated Case
United States Of America Published Application 20190276842 09/12/2019 2017-016
European Patent Office Published Application 3523426 A0 08/14/2019 2017-016
China Published Application CN110023494A 07/16/2019 2017-016
India Published Application 28/2019 07/12/2019 2017-016
United Kingdom Published Application 2569733 06/26/2019 2017-016
Brazil Published Application 2529 06/25/2019 2017-016
United States Of America Published Application 20180346927 12/06/2018 2017-016
Australia Published Application WO 2018/064371 04/05/2018 2017-016
Canada Published Application WO 2018/064371 04/05/2018 2017-016
Eurasian Patent Office Published Application WO 2018/064371 04/05/2018 2017-016
Israel Published Application WO 2018/064371 04/05/2018 2017-016
Japan Published Application WO 2018/064371 04/05/2018 2017-016
Rep Of Korea Published Application WO 2018/064371 04/05/2018 2017-016
Mexico Published Application WO 2018/064371 04/05/2018 2017-016
New Zealand Published Application WO 2018/064371 04/05/2018 2017-016
Saudi Arabia Published Application WO 2018/064371 04/05/2018 2017-016
Singapore Published Application WO 2018/064371 04/05/2018 2017-016
South Africa Published Application WO 2018/064371 04/05/2018 2017-016
Patent Cooperation Treaty Published Application WO2018064371 04/05/2018 2017-016
 

Additional Patents Pending

Brief Description


The CRISPR-Cas system is now understood to confer bacteria and archaea with acquired immunity against phage and viruses. CRISPR-Cas systems consist of Cas proteins, which are involved in acquisition, targeting and cleavage of foreign DNA or RNA, and a CRISPR array, which includes direct repeats flanking short spacer sequences that guide Cas proteins to their targets.  Class 2 CRISPR-Cas are streamlined versions in which a single Cas protein bound to RNA is responsible for binding to and cleavage of a targeted sequence. The programmable nature of these minimal systems has facilitated their use as a versatile technology that is revolutionizing the field of genome manipulation.  Current CRISPR Cas technologies are based on systems from cultured bacteria, leaving untapped the vast majority of organisms that have not been isolated.  There is a need in the art for additional Class 2 CRISPR/Cas systems (e.g., Cas protein plus guide RNA combinations).

 

UC Berkeley researchers discovered a new type of Cas protein, CasX, from groundwater samples. CasX is short compared to previously identified CRISPR-Cas endonucleases, and thus use of this protein as an alternative provides the advantage that the nucleotide sequence encoding the protein is relatively short.  CasX utilizes a tracrRNA and a guide RNA to perform double stranded cleavage of DNA. The researchers introduced CRISPR-CasX into E. coli, finding that they could block genetic material introduced into the cell.  Further research results indicated that CRISPR-CasX operates in a manner analogous to CRISPR-Cas9, but utilizing an entirely distinct protein architecture containing different catalytic domains.   CasX is also expected to function under different conditions (e.g., temperature) given the environment of the organisms that CasX was expressed in.  Similar to CRISPR Cas9, CasX enzymes are expected to have a wide variety of applications in genome editing and nucleic acid manipulation. 



Suggested uses


  • Genome editing
  • Genetic engineering
  • Gene therapy
  • Research tools (e.g., high-throughput screening of gene functions in cell lines and in vivo)
  • Creation of transgenic animal models
  • Genomic imaging

 

Advantages


  • Functions under different conditions than currently used CRISPR-Cas proteins (e.g., lower temperatures)
  • Nucleotide sequence encoding the CasX protein is short, therefore it's especially useful in situations that employ a viral vector (e.g., an AAV vector), for delivery to a cell such as a eukaryotic cell 

 

Publications

CasX enzymes comprise a distinct family of RNA-guided genome editors: Jun-Jie Liu, Natalia Orlova, Benjamin L. Oakes, Enbo Ma, Hannah B. Spinner, Katherine L. M. Baney, Jonathan Chuck, Dan Tan, Gavin J. Knott, Lucas B. Harrington, Basem Al-Shayeb, Alexander Wagner, Julian Brötzmann, Brett T. Staahl, Kian L. Taylor, John Desmarais, Eva Nogales & Jennifer A. Doudna, Nature, volume 566, pages218–223 (2019) 

New CRISPR–Cas systems from uncultivated microbes 

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Inventors

  • Doudna, Jennifer A.

Other Information

Keywords

CRISPR, gene editing, genome, gene therapy, cell biology, CasX

Categorized As