Ludwig-Maximilian-Universitat, BioMedical Center 2 articles published in JoVE Genetics A Customizable Protocol for String Assembly gRNA Cloning (STAgR) Christopher T. Breunig1,2, Andrea M. Neuner1,2, Jessica Giehrl-Schwab3, Wolfgang Wurst3, Magdalena Götz2,4, Stefan H. Stricker1,2 1MCN Junior Research Group, Munich Center for Neurosciences, Ludwig Maximilian Universitat, BioMedical Center, 2Institute of Stem Cell Research, Helmholtz Zentrum, German Research Center for Environmental Health, 3Institute of Developmental Genetics, Helmholtz Zentrum, German Research Center for Environmental Health, 4Physiological Genomics, Ludwig Maximilian Universitat, BioMedical Center Here, we present string assembly gRNA cloning (STAgR), a method to easily multiplex gRNA vectors for CRISPR/Cas9 approaches. STAgR makes gRNA multiplexing simple, efficient and customizable. Genetics A Universal Protocol for Large-scale gRNA Library Production from any DNA Source Anna Köferle1, Stefan H. Stricker1,2 1MCN Junior Research Group, Munich Center for Neurosciences, Ludwig-Maximilian-Universität, BioMedical Center, 2Epigenetic Engineering, Institute of Stem Cell Research, Helmholtz Zentrum, German Research Center for Environmental Health Methods for generating large-scale gRNA libraries should be simple, efficient and cost-effective. We describe a protocol for the production of gRNA libraries based on enzymatic digestion of target DNA. This method, CORALINA (comprehensive gRNA library generation through controlled nuclease activity) presents an alternative to costly custom oligonucleotide synthesis.
