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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Miyaoka, Yuichiro Chan, Amanda H. Conklin, Bruce R. |
| Description | Author Affiliation: Miyaoka Y ( Gladstone Institute of Cardiovascular Disease, San Francisco, California 94158); Chan AH ( Gladstone Institute of Cardiovascular Disease, San Francisco, California 94158); Conklin BR ( Gladstone Institute of Cardiovascular Disease, San Francisco, California 94158) |
| Abstract | The detection of genome editing is critical in evaluating genome-editing tools or conditions, but it is not an easy task to detect genome-editing events-especially single-nucleotide substitutions-without a surrogate marker. Here we introduce a procedure that significantly contributes to the advancement of genome-editing technologies. It uses droplet digital polymerase chain reaction (ddPCR) and allele-specific hydrolysis probes to detect single-nucleotide substitutions generated by genome editing (via homology-directed repair, or HDR). HDR events that introduce substitutions using donor DNA are generally infrequent, even with genome-editing tools, and the outcome is only one base pair difference in 3 billion base pairs of the human genome. This task is particularly difficult in induced pluripotent stem (iPS) cells, in which editing events can be very rare. Therefore, the technological advances described here have implications for therapeutic genome editing and experimental approaches to disease modeling with iPS cells. |
| File Format | HTM / HTML |
| ISSN | 19403402 |
| Issue Number | 8 |
| Journal | Cold Spring Harbor Protocols |
| Volume Number | 2016 |
| e-ISSN | 15596095 |
| Language | English |
| Publisher | Cold Spring Harbor Laboratory Press |
| Publisher Date | 2016-08-01 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Clinical Laboratory Techniques |
| Content Type | Text |
| Resource Type | Article |
| Subject | Biochemistry, Genetics and Molecular Biology |
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