One caveat to these experiments is that the mutants used may not completely eliminate the function of the cell.
Use of perforin-deficient pfpCTLs and Fas-resistant CT26 tumor cells will eliminate the function of perforin and Fas/FasL effector mechanisms of the CTLs, thus reducing the CTL cytotoxicity background for optimal TRAIL function evaluation.
CBX also blocked the formation of primordial follicles possibly by inhibiting the expression of PAR6 for that eliminating the function of PAR6 by anti-PAR6 antibody and RNAi produced similar phenotypes of reducing the number of primordial follicles.
Therefore, we eliminated the function of FLRT1 and FLRT3 proteins by either knocking down their expression with morpholinos or by knocking down the expression of RND1, the downstream effector of FLRT proteins, as done previously [2].
Consequently, mutations that drastically reduce or eliminate the function of most of the nervous system are easily isolated.
We took advantage of an existing transposon insertion into the peel-1 gene, which is predicted to eliminate the function of peel-1.
If we eliminate the function of peel-1, the strain will be compatible with any other strain, including the Hawaii strain.
We also show that reducing SV release by eliminating the function of UNC-13L C2A domain ameliorates behavioral deficits in a C. elegans model for epileptic seizure.
Since RNAi effectively reduces but does not eliminate the function of LNKN-1, we interpret the similar but stronger phenotype of the mutant to indicate that the mutant is in fact a loss-of-function allele.
Note that only in one of these eight cases did the well-conserved amino acid itself change, therefore this bias would appear to have more to do with modulating rather than eliminating the function of these conserved residues.
