Fig. 4
CRISPR-dCas13b-ADAR2DD mediated knockdown of K48-polyubiquitination in the BLA impairs contextual fear memory in female, but not male, rats. Young adult male and female rats (n = 8 per group per sex) received a bilateral infusion of either gRNA targeting K48 and dCas13b-ADAR2DD plasmids (K48 + dCas13; Treatment) or dCas13b-ADAR2DD plasmid alone (Control) into the BLA. Twenty-eight days later, rats were trained to contextual fear conditioning and tested the following day. A During contextual fear conditioning, we found a main effect for Time in (F2.95,82.61 = 91.21, p < 0.0001) and Sex (F1,28 = 6.572, p = 0.0160), but there was no main effect for Treatment (F1,28 = 1.242, p = 0.2746), nor was there a significant interaction of Time X Sex (F4,112 = 0.4820, p = 0.7489), Time X Treatment (F4,112 = 1.464, p = 0.2178), Sex X Treatment (F1,28 = 2.742, p = 0.1089), or Time X Sex X Treatment (F4,112 = 1.412, p = 0.2346). B During testing, there was no main effect for Treatment (F1,28 = 2.866, p = 0.1016), but there was a main effect for Sex (F1,28 = 34.40, p < 0.0001) and there was a significant interaction of Treatment X Sex (F1,28 = 6.358, p = 0.0177). Bonferroni post hoc analysis determined that K48 + dCas13 males had no difference in performance compared to control males (p > 0.9999), but K48 + dCas13 females had significantly impaired memory compared to control females (p = 0.0118), indicating that the K48-polyubiquitin knockdown only impaired memory retention in females. *p < 0.05