CRISPR/Cas9-Directed genome editing of cultured cells. Yang, L., Yang, J. L, Byrne, S., Pan, J., & Church, G. M Current protocols in molecular biology / edited by Frederick M. Ausubel ... [et al.], 107(July):31.1.1–31.1.17, January, 2014. ISBN: 0471142727![CRISPR/Cas9-Directed genome editing of cultured cells. [link]](https://bibbase.org/img/filetypes/link.svg "link")
Paper doi abstract bibtex Human genome engineering has been transformed by the introduction of the CRISPR (clustered regularly interspaced short palindromic repeats)/Cas (CRISPR-associated) system found in most bacteria and archaea. Type II CRISPR/Cas systems have been engineered to induce RNA-guided genome editing in human cells, where small RNAs function together with Cas9 nucleases for sequence-specific cleavage of target sequences. Here we describe the protocol for Cas9-mediated human genome engineering, including construct building and transfection methods necessary for delivering Cas9 and guide RNA (gRNA) into human-induced pluripotent stem cells (hiPSCs) and HEK293 cells. Following genome editing, we also describe methods to assess genome editing efficiency using next-generation sequencing and isolate monoclonal hiPSCs with the desired modifications for downstream applications. Curr. Protoc. Mol. Biol. 107:31.1.1-31.1.17. © 2014 by John Wiley & Sons, Inc.
@article{Yang2014,
title = {{CRISPR}/{Cas9}-{Directed} genome editing of cultured cells.},
volume = {107},
issn = {1934-3647},
url = {http://www.ncbi.nlm.nih.gov/pubmed/24984853},
doi = {10.1002/0471142727.mb3101s107},
abstract = {Human genome engineering has been transformed by the introduction of the CRISPR (clustered regularly interspaced short palindromic repeats)/Cas (CRISPR-associated) system found in most bacteria and archaea. Type II CRISPR/Cas systems have been engineered to induce RNA-guided genome editing in human cells, where small RNAs function together with Cas9 nucleases for sequence-specific cleavage of target sequences. Here we describe the protocol for Cas9-mediated human genome engineering, including construct building and transfection methods necessary for delivering Cas9 and guide RNA (gRNA) into human-induced pluripotent stem cells (hiPSCs) and HEK293 cells. Following genome editing, we also describe methods to assess genome editing efficiency using next-generation sequencing and isolate monoclonal hiPSCs with the desired modifications for downstream applications. Curr. Protoc. Mol. Biol. 107:31.1.1-31.1.17. © 2014 by John Wiley \& Sons, Inc.},
number = {July},
journal = {Current protocols in molecular biology / edited by Frederick M. Ausubel ... [et al.]},
author = {Yang, Luhan and Yang, Joyce L and Byrne, Susan and Pan, Joshua and Church, George M},
month = jan,
year = {2014},
pmid = {24984853},
note = {ISBN: 0471142727},
keywords = {\#nosource, genome engineering r crispr, r human stem cells},
pages = {31.1.1--31.1.17},
}
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