CRISPR-Display

CRISPR-Display (CRISP-Disp) is a modification of the CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats) system for genome editing. The CRISPR/Cas9 system uses a short guide RNA (sgRNA) sequence to direct a Streptococcus pyogenes Cas9 nuclease, acting as a programmable DNA binding protein, to cleave DNA at a site of interest.[1][2]

CRISPR-Display, in contrast, uses a nuclease deficient Cas9 (dCas9) and an engineered sgRNA with aptameric accessory RNA domains, ranging from 100bp to 5kb, outside of the normal complementary targeting sequence.[3] The accessory RNA domains can be functional domains, such as long non-coding RNAs (lncRNAs), protein-binding motifs, or epitope tags for immunochemistry. This allows for investigation of the functionality of certain lncRNAs, and targeting of ribonucleoprotein (RNP) complexes to genomic loci.

CRISPR-Display was first published in Nature Methods in July 2015, and developed by David M. Shechner, Ezgi Hacisuleyman, Scott T. Younger and John Rinn at Harvard University and Massachusetts Institute of Technology (MIT), USA.

  1. ^ Jinek, Martin; Chylinski, Krzysztof; Fonfara, Ines; Hauer, Michael; Doudna, Jennifer A.; Charpentier, Emmanuelle (2012-08-17). "A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity". Science. 337 (6096): 816–821. Bibcode:2012Sci...337..816J. doi:10.1126/science.1225829. ISSN 1095-9203. PMC 6286148. PMID 22745249.
  2. ^ Cong, Le; Ran, F. Ann; Cox, David; Lin, Shuailiang; Barretto, Robert; Habib, Naomi; Hsu, Patrick D.; Wu, Xuebing; Jiang, Wenyan (2013-02-15). "Multiplex genome engineering using CRISPR/Cas systems". Science. 339 (6121): 819–823. Bibcode:2013Sci...339..819C. doi:10.1126/science.1231143. ISSN 1095-9203. PMC 3795411. PMID 23287718.
  3. ^ Shechner, David M.; Hacisuleyman, Ezgi; Younger, Scott T.; Rinn, John L. (2015-07-01). "Multiplexable, locus-specific targeting of long RNAs with CRISPR-Display". Nature Methods. 12 (7): 664–670. doi:10.1038/nmeth.3433. ISSN 1548-7105. PMC 4821475. PMID 26030444.

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