Investigating the role of SNU66 in 5' splice site maintenance

CryoEM modeling of the human pre-B complex suggests an interaction between SNU66(H734) and SNRP27(M141) at the base of the ACAGAGA box of U6. We had previously shown in C. elegans that SNRP-27(M141T) mutants alter 5'ss usage. Here we generate new alleles by CRISPR in snu-66 at the corresponding H765 location in C. elegans (H765G and H765L) and demonstrate that they have similar effects as SNRP-27(M141T) mutants on 5'ss selection globally. Alternative splicing analysis of snu-66(H765G), snu-66(H765L) and snrp-27(M141T) mutant strains at larval stage L3 compared to N2 wild type