It has been clearly shown that it is possible to substantially improve the quality of sgRNA libraries with just a few changes - HE insertion1 and AT inversion2 - in the constant 3′-sequence of sgRNAs. Libraries with this modified sgRNA structure knock out target genes more quickly and effectively ...Learn more
Cellecta routinely uses the HEAT-modified sgRNA design in its constructs and libraries.
1 Addition of a 5 nucleotide extension (HE) to the stem of a stem-loop structure making it more stable and accessible to the Cas9 protein.
2 Place swap of an adenine (A) and thymine (T) to remove a transcription terminator site.