Part:BBa_K2200012

_NOTOC__ the miRNA was designed to target and downregulate the expression of SAMMSON gene.

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Usage and Biology

A microRNA (miRNA) is a small non-coding RNA molecules (about 22-25nt) that works in RNA silencing and post-transcriptional regulation of gene expression. [1] It disturbs the expression of genes with complementary sequences by binding and cleaving its mRNA and thus prevents translation.

Our microRNA is designed to specially bind to Survival Associated Mitochondrial Melanoma Specific Oncogenic Non-Coding RNA (SAMMSON) and regulate the expression of CRISPR/Cas9 system. We also added an amiRNA binding site complementary to our artificial microRNA after the Cas9 protein.

In normal cells, the microRNA binds to the amiRNA binding site, and the expression of Cas9 is down-regulated. In melanoma cells, however, the microRNA binds to the SAMMSON RNAs so it has no effect on the expression of CRISPR/Cas9. As a result, the expression of CRISPR/Cas9 system can be regulated. (More information, http://2017.igem.org/Team:Shenzhen_SFLS/Design)

The miRNA was designed base on the human microRNA 30a (sequence found in NCBI, https://www.ncbi.nlm.nih.gov/gene/407029) and BLOCK-iT™ RNAi Designer was used to design the microRNA. (http://rnaidesigner.thermofisher.com/rnaiexpress/design.do.)

Sequence and Features

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]

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Reference

[1] microRNA, wikipedia (https://en.wikipedia.org/wiki/MicroRNA)

[2] Leucci E, Vendramin R, Spinazzi M, et al. Melanoma addiction to the long non-coding RNA SAMMSON. Nature, 2016, 531(7595):518-522.