Part:BBa_K2644128
GGG+crRNA pAcGFP1-N2
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
This biological part is specially designed to target a GFP plasma. The principle behind this is that CRISPR-Cas12a (Cpf1) proteins are RNA-guided enzymes that bind and cut DNA as components of bacterial adaptive immune systems like CRISPR-Cas9. We designed the crRNA which is able to combine with the Cas12a protein and recognize the target DNA. According to this principle, we designed a crRNA that matched with one fragment of GFP plasma., which brings Cas12a to its target, specifically cleaves the GFP plasma. Through this method, we are theoretically capable of targeting any single base mutation on ctDNA which are dispersed in people’s blood.
Cleaving experiment
Figure 1.Result of sequence-specific plasmid cleavage. Line 1, plasmid GFP. Line 2, cleavage plasmid with BamH1 enzyme. Line 3, cleavage plasmid with Fncas12a,crRNA-1 and crRNA-2. Line 4, cleavage plasmid with Fncas12a and crRNA-1. Line 5, cleavage plasmid with Fncas12a and crRNA-2.
Figure 2.Result of specific-sequence cleavage after Optimization. Line 1, GFP plasmid. Line 2, BamH1 cleavage. Line 3, cleavage according to: crRNA：FnCas12a：plasmid = 10:10:1. Line 4, cleavage according to: crRNA：FnCas12a：Plasmid = 10:10:1.
| None |