Additional file 1 of Engineered circular guide RNAs boost CRISPR/Cas12a- and CRISPR/Cas13d-based DNA and RNA editing

Additional file 1: Fig. S1 Establishment of the dLbCas12a-p300knock-in HEK293T cell line. Fig. S2 Circular gRNAs increase thetranscription efficiency of LbCas12a-based activators. Fig. S3 Establishment of the LbCas12a knock-in HEK293Tcell line and off-target analysis of LbCas12a with different gRNAs. Fig. S4 Circular gRNAs improve the DNA cleavage efficiency of LbCas12ain MCF7 cells. Fig. S5 comparison between cgRNA with other extendedstructuralized gRNAs. Fig. S6 Circular gRNAs improve the gene expressionor DNA cleavage of AsCas12a-based effectors. Fig. S7 Multiplexedgene activation and cleavage guided by cgRNAs. Fig. S8 Efficient andspecific RNA cleavage activity of CasRX with cgRNA. Fig. S9 Thetrans-cleavage activity of CasRx–mediated exogenous transcripts degradation. Fig.S10 The trans-cleavage activity of CasRx–mediated endogenous transcriptsdegradation. Fig. S11 The structures of circular gRNAs for LbCas12awith different linkers targeting IL1RN predicted by mFold. Fig. S12The structures of circular gRNAs for LbCas12a with different linkerstargeting IL1RN predicted by mFold. Fig. S13 The structures ofcircular gRNAs for CasRx with different linkers targeting STAT3predicted by mFold.

附加文件1：
图S1 dLbCas12a-p300敲入型HEK293T细胞系的构建；
图S2 环状gRNA（circular gRNAs）可提升基于LbCas12a的激活效应物的转录效率；
图S3 LbCas12a敲入型HEK293T细胞系的构建，以及不同gRNAs介导的LbCas12a脱靶效应分析；
图S4 环状gRNA可提升MCF7细胞中LbCas12a的DNA切割效率；
图S5 环状gRNA与其他延伸结构化gRNA的对比分析；
图S6 环状gRNA可提升基于AsCas12a的效应物的基因表达或DNA切割能力；
图S7 环状gRNA介导的多重基因激活与切割实验；
图S8 结合环状gRNA的CasRX展现出高效且特异性的RNA切割活性；
图S9 CasRx介导的外源转录本降解的反式切割活性；
图S10 CasRx介导的内源转录本降解的反式切割活性；
图S11 经mFold预测的、靶向IL1RN且带有不同连接序列的LbCas12a环状gRNA的二级结构；
图S12 经mFold预测的、靶向IL1RN且带有不同连接序列的LbCas12a环状gRNA的二级结构；
图S13 经mFold预测的、靶向STAT3且带有不同连接序列的CasRx环状gRNA的二级结构。