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. Author manuscript; available in PMC: 2024 Feb 23.
Published in final edited form as: GEN Biotechnol. 2022 Jun 14;1(3):271–284. doi: 10.1089/genbio.2022.0003

FIG 1.

FIG 1.

New Cas12a NLS variants demonstrate improved activity in transformed cell lines. (A) Schematic representation of the prior designs for Asp/LbaCas12a (2xNLS-NLP-SV40) and enAspCas12a (1xNLS-NLP) in addition to the new Asp/enAsp/LbaCas12a NLS framework variants described in this study.13,14,27,40 (B) Quantification of editing efficiency by Cas12a RNP targeting AAVS1 site 1 in HEK293T cells at 5 pmol of protein:crRNA complex when delivered by nucleofection. (C) Quantification of editing efficiency by Cas12a RNP targeting AAVS1 site 1 in Jurkat cells at 5 pmol of protein:crRNA complex when delivered by nucleofection. (D) Quantification of editing efficiency by Cas12a RNP targeting AAVS1 site 1 in K562 cells at 5 pmol of protein:crRNA complex when delivered by nucleofection. Results were obtained from three independent experiments and presented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 by one-way ANOVA with Tukey’s multiple comparisons test between each NLS variant. Cas12a ortholog is denoted by color from 1A. Each NLS architecture is denoted by a number from 1A.