Part:BBa_K2908333

miR141-BD

When this part binding with miR141, it can inhibit the translation of mRNA which include the same sequence.This part is designed by assembling several complemental sequence with miR141 together and synthesized by company.It is used as a target aimed by miR141 to block the gene circuit in normal cells.

Usage and Biology

- √ miRNA-BD (microRNA binding sites) Design
We designed, constructed, and sequence validated a miRNA sensor library containing all 620 sequences of mature human miRNAs designated as high confidence in miRBase 2124. Our library enables high-information-content screening of miRNA activity in cells and also serves as a source for sequence-validated templates of miRNA targets when building multi-input sensors. We synthesized miRNA target site sets bearing four repeats of the sequence perfectly complementary to the miRNA and inserted them into the 3′ UTR of a reporter construct.
It’s worth mentioning that, the design of this part is instructed by the 2019 team SYSU_CHINA (Detailed information of collaboration can be seen in the Team/Collaboration page in our wiki. Address: http://2019.igem.org/Team:CSU_CHINA/Collaborations)
- Construction: miRNA-BDs were constructed bearing four repeats of miR-21-5p in the 3’ UTR, or 5’ UTR. Since these miRNA-BDs do not have ATGs, several ATGs were added between the miRNA target sites in some sensors. Extra bases were added to separate the ATGs by a number of bases divisible by three. The distance between the last ATG and the true reporter start codon was either divisible by three or not .

Figure: The flow cytometry picture shows that miRNA-BD has approximately 30% efficiency inhibiting the endogenous miR-141

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Unknown
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]