Efficient Cloning and Sequence Validation of Repetitive and High GC-Content Short Hairpin RNAs

Sequencing-guided design of genetically encoded small RNAs targeting CAG repeats for selective inhibition of mutant huntingtin

“…To generate small RNA expression vectors, we cloned small RNA duplexes ( Table S1 ) into a U6 promoter-driven lentiviral vector expressing a separate EGFP-puromycin reporter 24 , 25 ( Figure S1 A). Small RNAs were expressed from two core constructs, an shRNA 24 ( Figure 1 B) or a miRNA ( Figure 1 C) scaffold.…”