| Chapter title |
Exploring CRISPR Interference by Transformation with Plasmid Mixtures: Identification of Target Interference Motifs in Escherichia coli
|
|---|---|
| Chapter number | 10 |
| Book title |
CRISPR
|
| Published in |
Methods in molecular biology, January 2015
|
| DOI | 10.1007/978-1-4939-2687-9_10 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-2686-2, 978-1-4939-2687-9
|
| Authors |
Cristóbal Almendros, Francisco J. M. Mojica, Almendros, Cristóbal, Mojica, Francisco J. M. |
| Editors |
Magnus Lundgren, Emmanuelle Charpentier, Peter C. Fineran |
| Abstract |
Plasmid transformation into a bacterial host harboring a functional CRISPR-Cas system targeting a sequence in the transforming molecule can be specifically hindered by CRISPR-mediated interference. In this case, measurements of transformation efficacy will provide an estimation of CRISPR activity. However, in order to standardize data of conventional assays (using a single plasmid in the input DNA sample), transformation efficiencies have to be compared to those obtained for a reference molecule in independent experiments. Here we describe the use of a transforming mixture of plasmids that includes the non-targeted vector as an internal reference to obtain normalized data which are unbiased by empirical variations. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 16 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 4 | 25% |
| Student > Master | 3 | 19% |
| Professor > Associate Professor | 2 | 13% |
| Student > Ph. D. Student | 1 | 6% |
| Unspecified | 1 | 6% |
| Other | 2 | 13% |
| Unknown | 3 | 19% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 7 | 44% |
| Agricultural and Biological Sciences | 5 | 31% |
| Unspecified | 1 | 6% |
| Medicine and Dentistry | 1 | 6% |
| Unknown | 2 | 13% |