Difference between revisions of "Part:BBa K3645011"

Revision as of 13:44, 17 October 2021

Target-AID (CBE)

Contains the full CDS of Target-AID, whose Cas9 part was replace with our lab's dCas9.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1099
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 4775
Illegal BamHI site found at 3378
Illegal XhoI site found at 4384
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Contribution From NNU-China 2021

Group: iGEM Team NNU-China 2021

Author: Yan Xu

Summary: Testing its gene editing efficiency in BL21 (DE3)

Characterization from iGEM21-NNU-China

Cytosine base editors (CBEs) enable targeted C•G-to-T•A conversions in genomic DNA, consisting of dSpCas9, CDA, and UGI [1]. It was first registered in 2020. In order to test the editing efficiency of this composite part, we construct the dual plasmid system based on the (BBa_K3645011). We selected the cadA, maeA, and maeB genes as the testing sites, and the related pTarget plasmids were constructed. Results showed that the (BBa_K3645011) can successfully work in the BL21 (DE3), and the editing efficiency of single gene editing, double genes editing and triple genes editing can reach 85%, 56% and 25%, respectively (Fig. 1). These results provide references for future iGEM teams to choose gene-editing tools in E.coli.

Fig.1 The gene editing efficiency of the part of dCas9-CDA-UGI.

Reference

[1] Zhao D, Li J, Li S, Xin X, Hu M, Price MA, Rosser SJ, Bi C, Zhang X. Glycosylase base editors enable C-to-A and C-to-G base changes. Nature Biotechnology. 2021; 39: 35–40.

@@ Line 40: / Line 40: @@
 :'''Fig.1''' The gene editing efficiency of the part of dCas9-CDA-UGI.
+<p><b><h2>Reference</h2></b></p>
+<p>[1] Zhao D, Li J, Li S, Xin X, Hu M, Price MA, Rosser SJ, Bi C, Zhang X. Glycosylase base editors enable C-to-A and C-to-G base changes. Nature Biotechnology. 2021; 39: 35–40.</p>
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