| Chapter title |
Enhancer RNAs
|
|---|---|
| Chapter number | 15 |
| Book title |
Enhancer RNAs
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-4035-6_15 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-4033-2, 978-1-4939-4035-6
|
| Authors |
Andrey, Guillaume, Spielmann, Malte, Guillaume Andrey, Malte Spielmann M.D., Ph.D., Malte Spielmann |
| Editors |
Ulf Andersson Ørom |
| Abstract |
Targeted mutagenesis is required to evaluate the function of DNA segments across the genome. In recent years the CRISPR/Cas9 technology has been widely used for functional genome studies and is partially replacing classical homologous recombination methods in different aspects. CRISPR/Cas9-derived tools indeed allow the production of a wide-range of engineered mutations: from point mutations to large chromosomal rearrangements such as deletions, duplications and inversions. Here we present a protocol to engineer Embryonic Stem Cells (ESC) with desired mutations using transfection of custom-made CRISPR/Cas9 vectors. These methods allow the in vivo modeling of congenital mutations and the functional interrogation of DNA sequences. |
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