title: New variants of CRISPR RNA-guided genome editing enzymes.
abstract: CRISPR-mediated genome editing using the Streptococcus pyogenes Cas9 enzyme is revolutionizing life science by providing new, precise, facile and high-throughput tools for genetic modification by the specific targeting of double-strand breaks in the genome of hosts. Plant biotechnologists have extensively used the S. pyogenes Cas9-based system since its inception in 2013. However, there are still some limitations to its even broader usage in plants. Major restrictions, especially in agricultural biotechnology, are the currently unclear regulatory status of plants modified with CRISPR/Cas9 and the lack of suitable delivery methods for some plant species. Solutions to these limitations could come in the form of new variants of genome editing enzymes that have recently been discovered and have already proved comparable to or even better in performance than S. pyogenes CRISPR/Cas9 in terms of precision and ease of delivery in mammal cells. Although some of them have already been tested in plants, most of them are less well known in the plant science community. In this review, we describe the following new enzyme systems engineered for genome editing, transcriptional regulation and cellular imaging-C2c2 from L. shahii; Cas9 from F. novicida, S. aureus, S. thermophiles, N. meningitidis; Cpf1 from F. novicida, Acidaminococcus and Lachnospiraceae; nickase, split, enhanced and other Cas9 variants from S. pyogenes; catalytically inactive SpCas9 linked to various nuclease or gene-regulating domains-with an emphasis on their advantages in comparison with the broadly used SpCas9. In addition, we discuss new possibilities they offer in plant biotechnology.
authors.affiliation: Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia.
authors.forename: Jana
authors.initials: J
authors.lastname: Murovec
authors.affiliation: Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia.
authors.forename: Žan
authors.initials: Ž
authors.lastname: Pirc
authors.affiliation: Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA, USA.
authors.forename: Bing
authors.initials: B
authors.lastname: Yang
mesh_terms: D001709:Biotechnology; D064113:CRISPR-Cas Systems / Q000235:genetics; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D000072669:Gene Editing / Q000379:methods*; D008954:Models, Biological; D030821:Plants, Genetically Modified / Q000235:genetics; D013297:Streptococcus pyogenes / Q000201:enzymology*
keywords: CRISPR effector C2c2; CRISPR/Cas9; Cpf1; genome editing; plant biotechnology; variants of Streptococcus pyogenes Cas9