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Fig. 3 | Genome Biology

Fig. 3

From: Multiplex enCas12a screens detect functional buffering among paralogs otherwise masked in monogenic Cas9 knockout screens

Fig. 3

Multiplex gene knockout with enCas12a. a Experimental design. EnCas12a crRNA dual-guide array design. Each construct targets two genes; each gene is targeted by 3 crRNA; each candidate paralog gene pair is targeted by 18 gene-gene constructs, with six gene-control constructs per gene (including both A-B and B-A orientations). b Evaluating synthetic lethality. Single mutant fitness (SMF) is the mean log fold change of control guides targeting a single gene. Expected double mutant fitness (DMF) is the sum of SMF. Observed DMF is the mean log fold change of dual-targeting constructs. Delta log fold change (dLFC) is the difference between observed and expected fold change. c QC plot showing separation of SMF of constructs targeting control essential and nonessential genes. d Scatter plot of all mirror constructs (same crRNA in A-B and B-A orientations) showing lack of positional effects. e SMF in this screen vs. gene BF in Avana data. f Distribution of dLFC scores for 403 gene pairs in each cell line

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