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. Author manuscript; available in PMC: 2024 Aug 17.
Published in final edited form as: Cell Chem Biol. 2023 Jun 29;30(8):879–892.e5. doi: 10.1016/j.chembiol.2023.06.007

Figure 1. Synthesis of biased sgRNA library template.

Figure 1.

A library based on the WT SpCas9 sgRNA sequence was synthesized with 5’ and 3’ constant regions (the Targeting Region and Loop 3, respectively, shown in black) and a 60 nucleotide partially randomized region (shown in gold). The random region was synthesized using phosphoramidite mixes such that each position in the randomized region had a 58% chance of being the WT nucleotide at that position and a 14% chance of being any of the other 3 nucleotides. The synthesized library was purified, extended, and transcribed as described in the Methods.