NEW YORK (GenomeWeb) – The Federation of European Biochemical Societies (FEBS) Journal has released a special issue focusing on how to use CRISPR/Cas9, comprising nine review papers by researchers including Harvard University's George Church and Norbert Perrimon, Francisco Mojica of Spain's Universidad de Alicante, and Memorial Sloan Kettering Cancer Center's Ralph Garippa.
The issue covers the discovery of CRISPR in prokaryotes, CRISPR screening, epigenetic editing, CRISPR in model organisms and disease modeling, guide RNA (gRNA) design, off-target effects, and a comparison with RNAi for therapeutic purposes.
"We hope these reviews, like the technology they describe, will be useful, grounded in scientific fact and providing guidance for discovery," John Doench, director of the Genetic Perturbation Platform at the Broad Institute, wrote in his introduction.
The reviews come as excitement continues to build for CRISPR-based technology and applications. The growth in papers mentioning the technology is exponential. But many researchers are still struggling with how to use it. "Even deeply ingrained and thoroughly vetted tools, such as restriction enzymes, antibodies, and Sanger sequencing, still sometimes fail in routine applications and require troubleshooting," Doench wrote. "Each review presents a level-headed assessment of how best to apply this technology, its strengths, and its limitations."
Mojica, who discovered CRISPR arrays in bacteria decades ago, co-authored a review with Francisco Rodriguez-Valera of the Universidad Miguel Hernandez to provide some background on the technology; Garippa, Linde Miles of Johns Hopkins University, and MSKCC's John Poirier described in vitro pooled knockout screening using CRISPR/Cas9; and George Church and several members of his lab discussed engineered Cas9 variants for epigenetic regulation in both gene expression activators and repressors.
Several reviews focused on the use of CRISPR for disease modeling and use in model organisms. MSKCC's Geulah Livshits and her colleagues wrote a review of using CRISPR/Cas9 to study gene function in vivo, focusing on disease modeling in mice; Northwestern University's Erik Andersen and Mostafa Zamanian wrote about using CRISPR/Cas9 in Caenorhabditis elegans to study tropical diseases; and the University of Pennsylvania's Kiran Musunuru and Jennie Lin described the considerations for using CRISPR to build cellular models of disease.
Several more reviews covered technical aspects of CRISPR. Harvard's Stephanie Mohr led a review of gRNA design for research applications, noting the importance of both bioinformatic prediction and experimental validation; Jiing-Juan Yee, of the City of Hope National Medical Center, discussed off-target effects; and Dirk Haussecker, a German consultant, compared CRISPR's ability to inhibit genes in a therapeutic setting to that of RNAi.