3TAT-DRBD |
GRKKRRQRRR GSH GRKKRRQRRR GSH GRKKRRQRRR GSHAGDLSAGFFMEELNTYRQKQGVVLKYQELPNSGPPHDRRFTFQVIIDGREFPEGEGRSKKEAKNAAAKLAVEILNKEKKA |
TAT, 10 amino acids, triple TAT expressed in series GSH as linkers 16 (underlined)DRBD, motif 1 of human PKR,36 80 amino acids, residues from 9-78 |
SA |
AEAGITGTWYNQLGSTFIVTAGADGALTGTYESAVGNAESRYVLTGRYDSAPATDGSGTALGWTVAWKNNYRNAHSATTWSGQYVGGAEARINTQWLLTSGTTEANAWKSTLVGHATFTKVKPSAA |
natural core SA 39126 amino acids, residues from 21-130, 13.3 KDaD128A mutation (underlined)40
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Nucleic acids sequence |
Comment |
sur-siRNA |
5′ GAAGCAGTTTGAAGAATTA dTdT 3′ |
Fluorescent siRNAs labeled with FAM or Cy7 at the 5′ end of the sense strand. |
A10 |
5′biotin-GGGAGGA(fC)GA(fU)G(fC)GGA(fU)(fC)AG(fC)(fC)A(fU)G(fU)(fU)(fU)A(fC)G(fU)(fC)A(fC)(fU)(fC)(fC)(fU)(fU)3′5′Cy7-GGGAGGA(fC)GA(fU)G(fC)GGA(fU)(fC)AG(fC)(fC)A(fU)G(fU)(fU)(fU)A(fC)G(fU)(fC)A(fC)(fU)(fC)(fC)(fU)(fU)-biotin 3′ |
To increase stability against nuclease degradation, the pyrimidines in the aptamer A10 were 2′-fluoro modified. |
A10-(sur-siRNA) chimera |
5′GGGAGGA(fC)GA(fU)G(fC)GGA(fU)(fC)AG(fC)(fC)A(fU)G(fU)(fU)(fU)A(fC)G(fU)(fC)A(fC)(fU)(fC)(fC)(fU)(fU)-GAAGCAGUUUGAAGAAUUA 3′ |
A10-(sur-siRNA antisense strand ) chimera After a simple annealing, the 2 strands form stable chimera.22
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5′FAM/Cy7-UAAUUCUUCAAACUGCUUC dTdT 3′ |
sur-siRNA sense strand The 5′ end of the sense strand was labeled with FAM or Cy7. |