Difference between revisions of "Part:BBa K2381012"

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	<partinfo>BBa_K2381012 short</partinfo>		<partinfo>BBa_K2381012 short</partinfo>
		+	<p>
		+	It’s a gRNA sequence containing 20bp targeting at <i>oriC</i> and a full-length gRNA scaffold. The sequence is central to our project and is related with all the system built this year.<br/>
		+	The targeting sequence of this part is acquired from xxxx. Its function is proved by OD measurement after cotransformed with plasmid containing dCas9.[[File:T--HZAU-China--Results_Figure1.gif|400px|thumb|left|]]. We can observe tight repression on cell growth. Further investigation is conducted by qPCR and flow cytometric.
		+	</p>
		+	<h3>References</h3>
−	It’s a gRNA plasmid which helps our CRISPR-System target to the MG1655 OriC sequence in order to achieve our expectation
−	this year. Its backbone is taken from iGEM Kit Plate named BBa_K165005, but the BsaI restriction site which was in Amp.
−	coding sequence has been deleted and a synonymous mutation has occurred by overlapping PCR. We used J23119 promoter, a
−	high-intensity promoter, to obtain a higher expression of gRNA than dCas9. The 21bp-sequence comes after is a kind of
−	sequence types taken from the article that is complementary to the Oric sequence and has a nice effect on our project. The
−	full length gRNA scaffold is also designed after consulting literatures.
−
−	<h3>Reference</h3>
−	<p>Mitchell R. O’Connell, Benjamin L. Oakes, Samuel H. Sternberg, Alexandra East-Seletsky, Matias Kaplan & Jennifer A. Doudna (2014). Programmable RNA recognition and cleavage by CRISPR/Cas9. Nat Commun. doi:10.1038/nature13769</p>
−	<p>Alexandra E. Briner, Paul D. Donohoue, Ahmed A. Gomaa, Kurt Selle, Euan M. Slorach, Christopher H. Nye,Rachel E. Haurwitz, Chase L. Beisel, Andrew P. May, and Rodolphe Barrangou (2014). Guide RNA Functional Modules Direct Cas9 Activity and Orthogonality. Molecular Cell 56, 333–339, October 23, 2014 ª2014 Elsevier Inc. 333</p>
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Revision as of 14:10, 1 November 2017

3'-OriC gRNA

It’s a gRNA sequence containing 20bp targeting at oriC and a full-length gRNA scaffold. The sequence is central to our project and is related with all the system built this year.
The targeting sequence of this part is acquired from xxxx. Its function is proved by OD measurement after cotransformed with plasmid containing dCas9.

. We can observe tight repression on cell growth. Further investigation is conducted by qPCR and flow cytometric.

References

Sequence and Features