Part:BBa_K4778003

sgRNA_D

Usage and Biology

Sequence：5‘-UAA UUU CUA CUA AGU GUA GAU GAU CGU UAC GCU AAC UAU GA-3’

Length: 41nt

BBa_K2200000,Sigle-guide RNA (sgRNA), as mentioned by iGEM17_Shenzhen_SFLS, is an artificial RNA designed to combine specific DNA sequences. It can bind to Cas12a protein and play a role in the cleaving of target dsDNA. It has the same function as wild-type gRNA in CRISPR/Cas12a system, but with higher efficiency. We keep using the sgRNA used in the CONAN system to specifically target the double-stranded assistant DNA we used in pairs. It has no effect on normal cells and genes.

Fig1，Detailed sequence of the gRNA used in CONAN. The 5’-handle contains 21nucleotides and the length of 3’-spacer is 20 nt.

Fig2, The LC-MS data of sgRNA_D.

Results

In our system, together with DNA components, the sgRNA_D of the new probe is mainly responsible for the conversion and amplification of nucleic acid signals. After cleavage of DSN enzyme, it is proved that sgRNA is released.

Fig3，Denaturing PAGE. Arrow denotes sgRNA

Reference

[1]Shi K, Xie S, Tian R, et al. A CRISPR-Cas autocatalysis-driven feedback amplification network for supersensitive DNA diagnostics. Sci Adv. 2021;7(5):eabc7802. Published 2021 Jan 27.

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]