title: Guide RNA engineering for versatile Cas9 functionality.
abstract: The Clustered Regularly Interspaced Short Palindromic Repeats system allows a single guide RNA (sgRNA) to direct a protein with combined helicase and nuclease activity to the DNA. Streptococcus pyogenes Cas9 (SpCas9), a CRISPR-associated protein, has revolutionized our ability to probe and edit the human genome in vitro and in vivo Arguably, the true modularity of the Cas9 platform is conferred through the ease of sgRNA programmability as well as the degree of modifications the sgRNA can tolerate without compromising its association with SpCas9 and function. In this review, we focus on the properties and recent engineering advances of the sgRNA component in Cas9-mediated genome targeting.
authors.affiliation: Department of Biological Sciences, The University of Texas at Dallas, Richardson, TX 75080, USA.|Center for Systems Biology, The University of Texas at Dallas, Richardson, TX 75080, USA.
authors.forename: Chance M
authors.initials: CM
authors.lastname: Nowak
authors.affiliation: Department of Biological Sciences, The University of Texas at Dallas, Richardson, TX 75080, USA.
authors.forename: Seth
authors.initials: S
authors.lastname: Lawson
authors.affiliation: Department of Biological Sciences, The University of Texas at Dallas, Richardson, TX 75080, USA.|Center for Systems Biology, The University of Texas at Dallas, Richardson, TX 75080, USA.
authors.forename: Megan
authors.initials: M
authors.lastname: Zerez
authors.affiliation: Department of Biological Sciences, The University of Texas at Dallas, Richardson, TX 75080, USA bleris@utdallas.edu.|Center for Systems Biology, The University of Texas at Dallas, Richardson, TX 75080, USA.|Bioengineering Department, The University of Texas at Dallas, Richardson, TX 75080, USA.
authors.forename: Leonidas
authors.initials: L
authors.lastname: Bleris
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D000076987:CRISPR-Associated Protein 9; D064113:CRISPR-Cas Systems; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004720:Endonucleases / Q000378:metabolism*; D000072669:Gene Editing; D018390:Gene Targeting; D005818:Genetic Engineering; D006801:Humans; D011485:Protein Binding; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D000094704:RNA, Guide, CRISPR-Cas Systems; D000076987:CRISPR-Associated Protein 9; C000606107:Cas9 endonuclease Streptococcus pyogenes; D004720:Endonucleases