A PCR-based strategy for cloning short hairpin sequences:“PCR SHAGging”
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Gregory Hannon, Cold spring harbor lab
Our overall approach is to use an RNA polymerase III promoter to drive
expression of encoded short hairpin RNA (shRNA). For this purpose we use the human
U6 snRNA promoter, maintaining the transcript initiating “G” nucleotide of the
U6snRNA transcript. There by, hairpin sequences will start with a “G”. Termination is
mediated by a run of Ts at the end of the hairpin.
