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Helmholtz Zentrum, German Research Center for Environmental Health

2 ARTICLES PUBLISHED IN JoVE

Genetics

A Universal Protocol for Large-scale gRNA Library Production from any DNA Source

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.

Genetics

A Customizable Protocol for String Assembly gRNA Cloning (STAgR)

Christopher T. Breunig^1,2, Andrea M. Neuner^1,2, Jessica Giehrl-Schwab³, Wolfgang Wurst³, Magdalena Götz^2,4, Stefan H. Stricker^1,2

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.