|
| Status |
Public on Mar 02, 2020 |
| Title |
Randomized PAM depletion of phylogenetically-diverse Cas12a nucleases |
| Organism |
synthetic construct |
| Experiment type |
Other
|
| Summary |
We report the PAMs of phylogenetically-diverse Cas12a nucleases using a cell-free TXTL-based PAM screen. By adding a 5N randomized PAM library and Cas12a-gRNA in vitro, recognized PAM sequences were cleaved, while non-recognized PAMs remained. By amplifying the non-cleaved DNA, we used next-generation sequencing to analyze the depletion of functional PAMs of these Cas12a nucleases.
|
| |
|
| Overall design |
Uncleaved DNA from the randomized PAM library was amplified using Illumina Nextera Indices
|
| |
|
| Contributor(s) |
Jacobsen T |
| Citation(s) |
32329776 |
| |
| Submission date |
Apr 26, 2019 |
| Last update date |
Jun 08, 2020 |
| Contact name |
Chase Beisel |
| E-mail(s) |
[email protected]
|
| Organization name |
North Carolina State University
|
| Department |
Chemical and Biomolecular Engineering
|
| Lab |
Partners II, Rm 2110
|
| Street address |
840 Main Campus Drive
|
| City |
Raleigh |
| State/province |
NC |
| ZIP/Postal code |
27606 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL17769 |
Illumina MiSeq (synthetic construct) |
|
| Samples (12)
|
| GSM3737076 |
Adurb193Cas12a - T gRNA [Adurb193_S1] |
| GSM3737077 |
Adurb193Cas12a - NT gRNA [Adurb193-NT_S2] |
| GSM3737078 |
Adurb336Cas12a - T gRNA [Adurb336_S3] |
|
| Relations |
| BioProject |
PRJNA539942 |
| SRA |
SRP193973 |
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