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Subcloning of any sgRNA via BbsI sites. Note that there is an improved version of this plasmid (Addgene 105866) that results in up to 10x greater efficiency
Transient expression in mammalian cells of an "INT" construct_bearing P4_P6 (internally appended with three PP7 stem-loops), targeting the GLuc reporter. U6 promoter
Transient expression of an "INT" construct [bearing six minimal Puf1 stem-loops; 'INT' design: Shechner et al., Nature Methods 2015], in mammalian cells, U6 promoter