Part:BBa_K5407004
sh-TEAD4-2 sequence
Usage and Biology
Concerned with the fact that siRNA that is easy to be degraded in the cell, the shRNA plasmid can be more stable and keep releasing the siRNA, which would downregulate the expression of the TEAD4 more effectively [1-2]. Then, we designed the sh-TEAD4-2 sequence linked to the pLKO.1 plasmid.
Desigh
According to each siRNA target sequence, two complementary DNA template single strands are designed, including the siRNA sense strand and the antisense strand. Two strands are connected by a loop structure of 6 nucleotides. And Age I and EcoR I enzyme cleavage sequence are designed at both ends of the template strand (Figure 1).
Forward oligo: 5’ CCGG—21bp sense—CTCGAG—21bp antisense—TTTTTG 3’
Reverse oligo: 5’ AATTCAAAAA—21bp sense—CTCGAG—21bp antisense 3’
Figure 1 Model of shRNA design
Reference
[1] Walhan, Alshaer,Hadil, Zureigat,Arwa, Al Karaki et al. siRNA: Mechanism of action, challenges, and therapeutic approaches.[J] .Eur J Pharmacol, 2021, 905: 0.
[2] P N, Pushparaj,J J, Aarthi,J, Manikandan et al. siRNA, miRNA, and shRNA: in vivo applications.[J] .J Dent Res, 2008, 87: 0.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 26
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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