We have performed the experiment suggested by the reviewers by introducing a LARP7 mutant Δ2A that was originally found in microsatellite instable gastric cancer and contains a deletion of 2 adenosines from a microsatellite repeat of 8 A's (nucleotides 1206 1213) in the LARP7 C-terminal region.
If Notch signalling was indeed affected by CLN3 overexpression, we reasoned that introducing a mutant allele of the Notch ligand, Delta, should enhance the effect.
In order to test the functional effect of the human specific amino acids changes, we first constructed two mutant hMCPH1s by introducing a backward mutation of changing the human-specific amino acid to an ancestral amino acid (Met96Thr) and introducing a random mutation (Met96Leu) at Site-96.
Using lentiviral vectors, we genetically modified EGFR function by introducing an inactive mutant of the receptor in EGFR-amplified tumors.
In future, a more prolonged rescue might be obtained in vivo by upregulating Gdnf/Ret signalling in Fras1 mutant mice by introducing a heterozygous Sprouty1 mutation, as used to rescue Itga8 mutants (22).
First, it is a labor-intensive method to identify the genes responsible for the mutant phenotype because it requires phenotype rescue by introducing a cDNA library into the mutant clones, the identification of rescued cells, and the identification of responsible cDNAs from the rescued cells.
To examine the cooperative role of the OSR family members in organ growth, we generated an osr2 argos-1 ARLi triple mutant and an argos-1 osr1 ARLi OSR2i quadruple mutant by introducing a pro35S OSR2 RNAi construct into argos-1 osr1 ARLi plants.
The first mutant was generated by introducing a tyrosine to phenylalanine mutation at amino acid 14 of the first extracellular domain of CX3CR1 (Y14F) [ 31].
We generated extracellular signal-regulated kinase 1/2 (ERK1/2) mutants by introducing a single amino-acid substitution in subdomain V of the catalytic domain and then examined the susceptibility of these mutants to PP1 derivatives originally designed as Src inhibitors.
The mobility of the loop was reduced by introducing a new disulphide bridge in the mutant D241C/D249C.
