Difference between revisions of "Part:BBa K2908444"

Latest revision as of 00:27, 22 October 2019

miR148b-BD

When binding with miR148b, it can inhibit the translation of mRNA which include the same sequence.This part is designed by assembling several complemental sequence with miR148b together and synthesized by company.It is used as a target aimed by miR148b 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 20% efficiency inhibiting the endogenous miR-148b

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 5: / Line 5: @@
 When binding with miR148b, it can inhibit the translation of mRNA which include the same sequence.This part is designed by assembling several complemental sequence with miR148b together and synthesized by company.It is used as a target aimed by miR148b to block the gene circuit in normal cells.
-https://static.igem.org/mediawiki/parts/6/67/T--CSU_CHINA--miR148b-BD.png
+https://2019.igem.org/wiki/images/f/fb/T--CSU_CHINA--2908444.jpg
@@ Line 13: / Line 13: @@
 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)<br/>
 -	Construction: <br/>
-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 .
+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 . <br/>
+<center>https://2019.igem.org/wiki/images/6/6b/T--CSU_CHINA--miR148b-BS_small.jpg</center><br/>
+<center>Figure: The flow cytometry picture shows that miRNA-BD has approximately 20% efficiency inhibiting the endogenous miR-148b</center>
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