| Chapter title |
An Unbiased Method for Detection of Genome-Wide Off-Target Effects in Cell Lines Treated with Zinc Finger Nucleases
|
|---|---|
| Chapter number | 23 |
| Book title |
Gene Correction
|
| Published in |
Methods in molecular biology, January 2014
|
| DOI | 10.1007/978-1-62703-761-7_23 |
| Pubmed ID | |
| Book ISBNs |
978-1-62703-760-0, 978-1-62703-761-7
|
| Authors |
Cory R. Lindsay, David B. Roth, Lindsay, Cory R., Roth, David B. |
| Abstract |
We describe a method for detecting and validating genomic aberrations arising from cell lines exposed to zinc finger nucleases (ZFNs), an important reagent used for targeted genome modifications. This method makes use of cloned cell lines, an approach that adds power when testing variables that may affect gene correction efficiency and evaluating potential side effects on a genome-wide scale. After cell treatment, the genomic DNA isolation method, as described, is ideal for high-resolution array comparative genomic hybridization (aCGH) and quantitative PCR. Guidelines for aCGH analysis and calling significant copy number variations (CNVs) for validation by qPCR are also discussed. Using this method, we describe a novel ZFN-associated chromosome 4 copy number variation (CNV) attributable to a predicted ZFN off-target cleavage site found within the CNV. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 6 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 2 | 33% |
| Professor | 1 | 17% |
| Student > Bachelor | 1 | 17% |
| Researcher | 1 | 17% |
| Unknown | 1 | 17% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 3 | 50% |
| Medicine and Dentistry | 2 | 33% |
| Unknown | 1 | 17% |