Difference between revisions of "Part:BBa K2200007:Design"
Awesome Zz (Talk | contribs) |
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===Design Notes=== | ===Design Notes=== | ||
<p>Our modeling results showed that our sgRNA sequence has high cleavage efficiency on the mutated BRAF gene, as well as a high risk of off-target effect. To avoid the off-target effect on normal cells, we tried hard to develop an regulate system for the CRISPR/Cas9 system. At first, we thought about simply promote the system by the mutant hTERT but found it is a weak promoter. We then considered enhancing the function of it. We focused on the NF-KB p65 protein. However, we should change our direction since we found an article with results and their design is exactly the same as ours. So we have to look for other directions (that is, our present regulate system).</p> | <p>Our modeling results showed that our sgRNA sequence has high cleavage efficiency on the mutated BRAF gene, as well as a high risk of off-target effect. To avoid the off-target effect on normal cells, we tried hard to develop an regulate system for the CRISPR/Cas9 system. At first, we thought about simply promote the system by the mutant hTERT but found it is a weak promoter. We then considered enhancing the function of it. We focused on the NF-KB p65 protein. However, we should change our direction since we found an article with results and their design is exactly the same as ours. So we have to look for other directions (that is, our present regulate system).</p> | ||
| − | <p>We thought the Gal4-P65 could still be an improvement for BBa_K1470002 although we didn’t utilize it in our final project.</p> | + | <p>We thought the Gal4-P65 could still be an improvement for BBa_K1470002(https://parts.igem.org/Part:BBa_K1470002) although we didn’t utilize it in our final project.</p> |
Revision as of 14:47, 1 November 2017
Gal4 transcription factor is a positive regulator.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 710
Illegal XhoI site found at 218 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 886
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 137
Design Notes
Our modeling results showed that our sgRNA sequence has high cleavage efficiency on the mutated BRAF gene, as well as a high risk of off-target effect. To avoid the off-target effect on normal cells, we tried hard to develop an regulate system for the CRISPR/Cas9 system. At first, we thought about simply promote the system by the mutant hTERT but found it is a weak promoter. We then considered enhancing the function of it. We focused on the NF-KB p65 protein. However, we should change our direction since we found an article with results and their design is exactly the same as ours. So we have to look for other directions (that is, our present regulate system).
We thought the Gal4-P65 could still be an improvement for BBa_K1470002(https://parts.igem.org/Part:BBa_K1470002) although we didn’t utilize it in our final project.
Source
It comes from Saccharomyces cerevisiae.
References
[1] Huang X, Zhuang C, Zhuang C, et al. An enhanced hTERT promoter-driven CRISPR/Cas9 system selectively inhibits the progression of bladder cancer cells. Molecular Biosystems, 2017, 13(9): 1713-1721.