Difference between revisions of "Part:BBa K2200006"

Revision as of 01:55, 13 October 2017

pU6+sgRNA+pHEf1A+hCas9

This part is consisted of Human U6 promoter, sgRNA that targets the specific BRAF sequence. With its high flexibility of being devised to target different tumor gene sequences, it has a great market upon innovating cancer treatments since it will revolutionize this market by personalizing the remedy.

Usage and Biology

The composite part is specially designed to target BRAF tumor DNA. The principle behind this “device” is that when viruses invade bacterias , the self-protection system of bacteria detect the viruses’ DNA. The bacteria produces two kinds of RNA, one of which contains a sequence that matches the DNA of invading viruses, and this sequence is regarded as guide RNA(also small guide RNA, sgRNA). These two RNAs form a complex with Cas9 protein. When sgRNA finds its target, Cas9( the DNA scissor) then cuts the target DNA sequence. In order to conduct this principle, which is called CRISPR-Cas9, we designed a sgRNA that matches our experimental BRAF cell DNA. This sgRNA then brings Cas9 to its target to cut off the mutant DNA. It then causes the death of those cancer cells. Using this method, we are theoretically capable of targeting any tumor cells. This kind of innovated and precise methodology can be adopted to track patients throughout their life without getting their normal cells into danger and within suppressing the dispersion of cancer cells. U6 promoter drives the expression of sgRNA and HEf1A promoter drives the expression of Cas9.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 5117
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 863
Illegal XhoI site found at 1262
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 997
Illegal NgoMIV site found at 4214
Illegal NgoMIV site found at 5123
Illegal AgeI site found at 375
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 5242
Illegal SapI.rc site found at 2633
Illegal SapI.rc site found at 2875

@@ Line 9: / Line 9: @@
 ===Usage and Biology===
 The composite part is specially designed to target BRAF tumor DNA.
-The principle behind this “device” is that when viruses invade bacterias , the self-protection system of bacteria detect the viruses’ DNA. The bacteria produces two kinds pf RNA, one of which contains a sequence that matches the DNA of invading viruses, and this sequence is regarded as guide RNA(also small guide RNA, sgRNA). These two RNAs form a complex with Cas9 protein. When sgRNA finds its target, Cas9( the DNA scissor) then cuts the target DNA sequence.
+The principle behind this “device” is that when viruses invade bacterias , the self-protection system of bacteria detect the viruses’ DNA. The bacteria produces two kinds of RNA, one of which contains a sequence that matches the DNA of invading viruses, and this sequence is regarded as guide RNA(also small guide RNA, sgRNA). These two RNAs form a complex with Cas9 protein. When sgRNA finds its target, Cas9( the DNA scissor) then cuts the target DNA sequence.
 In order to conduct this principle, which is called CRISPR-Cas9, we designed a sgRNA that matches our experimental BRAF cell DNA. This sgRNA then brings Cas9 to its target to cut off the mutant DNA. It then causes the death of those cancer cells. Using this method, we are theoretically capable of targeting any tumor cells. This kind of innovated and precise methodology can be adopted to track patients throughout their life without getting their normal cells into danger and within suppressing the dispersion of cancer cells.
 U6 promoter drives the expression of sgRNA and HEf1A promoter drives the expression of Cas9.