Fig. 2

ccdc141 crispants in zebrafish displayed abnormal LR patterning. A: The schematic diagram documenting the process and guide RNAs used for generating ccdc141 crispants. B: Cardiac looping of zebrafish at 48 hpf is presented in the Tg(myl7:RFP) line in the ventral view. Normal D-loop, abnormal N-loop, and L-loop are shown. C: The quantification of cardiac looping (percentage). Cas9 control/crispants/crispants + mRNA, N = 3; n = 164/213/196; ????, P < 0.0001; two-sided Fisher's exact test for crispants with control and crispants + mRNA with control. D: Double WISH for myl7 (heart) and ins (pancreas) at 72 hpf. SA indicates any intermediate state between SS and SI. E: The percentage of abnormal organ situs with crispants. Control vs. crispants, N = 3; n = 63/107. ????, P < 0.0001; two-sided Fisher's exact test for crispants with control. F: Representative images of normal zebrafish (above), pericardial edema (middle), and curved tail (bottom) at 48 hpf. G and H: pericardial edema and curved tail of zebrafish (percentage). N = 6; ????, P < 0.0001; ??, P < 0.01; ?, P < 0.05; ns, not significant; two-sided Fisher's exact. WT, wild-type; R, right; L, left; SA, situs ambiguous; SS, situs solitus; SI, situs inversus; hpf, hours post fertilization.