Difference between revisions of "Part:BBa K3286041"

Latest revision as of 08:47, 17 October 2019

F. novicida Cpf1 (Cas12a) CRISPR array

Francisella novicida Cpf1 (Cas12a) CRISPR array .

The Cpf1 (Cas12a) CRISPR array of F. novicida, with two repeats and a scrambled (non-targeting) spacer.

New spacers can be generated by BbsI digestion of the CRISPR array and ligation of two annealed oligo's (with 5'-AGAT... and 5'-AGAC overhangs). By doing so, we can generate very fast, cheap and reliable a library of CRISPR arrays*.

(*We have observed that insertion of two spacers (with a repeat in the middle) is also possible with the use of ultramers.)

Zetsche, B., Gootenberg, J. S., Abudayyeh, O. O., Slaymaker, I. M., Makarova, K. S., Essletzbichler, P., … Zhang, F. (2015). Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell, 163(3), 759–771. https://doi.org/10.1016/j.cell.2015.09.038

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 10: / Line 10: @@
 By doing so, we can generate very fast, cheap and reliable a library of CRISPR arrays*.
-    *We have observed that insertion of two spacers (with a repeat in the middle) is also possible with the use of ultramers.
+(*We have observed that insertion of two spacers (with a repeat in the middle) is also possible with the use of ultramers.)
+<p>
+</p>
+<ol>
+<li>Zetsche, B., Gootenberg, J. S., Abudayyeh, O. O., Slaymaker, I. M., Makarova, K. S., Essletzbichler, P., … Zhang, F. (2015). Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell, 163(3), 759–771. https://doi.org/10.1016/j.cell.2015.09.038</li>
+</ol>
 <!-- Add more about the biology of this part here
 ===Usage and Biology===
 <!-- -->