Review Article

Variables and Strategies in Development of Therapeutic Post-Transcriptional Gene Silencing Agents

Table 3

Construction of allelic variant genes for combined knockdown reconstitute therapies.

Allelic variants for HhRz Therapeutics

5′ UT Target Site

5′…CCUGAGUGGCUGAGCUC AGGCCUU (5′ UT target site CUC )
5′…CCUGAGUGGCUGAGCUG AGGCCUU (aWT variant mRNA, CUG cannot be cleaved)

Coding V230 region
5′…CUC GUC UUC ACC GUC  AAG GAG GCC…3′(Coding region GUC )
L226 V227 F228 T229 V230 K231 E232 A233 (Amino acid triplets)
5′…CUC GUC UUC ACC GUG AAG GAG GCC…3′( WT variant mRNA,
L226 V227 F228 T229 V230 K231 E232 A233GUG cannot be cleaved)
Single letter amino acid codes are used.

Coding F293 region
5′…AUC CCA GCG UU C UUU GCC AAG AGC…3′(Coding region GUU )
I290 P291 A292 F293 F294 A295 K296 S297
GUU cleavage site occurs within the F293 codon rather than cutting at the end of a codon.
5′…AUC CCA GCG UGC UUU GCC AAG AGC…3′( F293C variant mRNA)
I290 P291 A292 C293 F294 A295 K296 S297
It is unclear whether or not the F293C mutation is an allelic variant WT or has a protein phenotype.

Allelic variant for RNAi therapeutics in F293 region
5′…AUC CCA GCG UUC UUU GCC AAG AGC…3′(Coding region RNAi site)
I290 P291 A292 F293 F294 A295 K296 S297
5′…AUA CCC GCA UUU UUC GCG AAA AGG…3′( WT variant mRNA)
I290 P291 A292 F293 F294 A295 K296 S297
aWT variant generated by codon degeneracy across the region of designed RNAi antisense annealing.