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@ARTICLE{Nasri:153073,
author = {M. Nasri and P. Mir$^*$ and B. Dannenmann and D. Amend and
T. Skroblyn and Y. Xu and K. Schulze-Osthoff$^*$ and M.
Klimiankou and K. Welte and J. Skokowa},
title = {{F}luorescent labeling of {CRISPR}/{C}as9 {RNP} for gene
knockout in {HSPC}s and i{PSC}s reveals an essential role
for {GADD}45b in stress response.},
journal = {Blood advances},
volume = {3},
number = {1},
issn = {2473-9537},
address = {Washington, DC},
publisher = {American Society of Hematology},
reportid = {DKFZ-2020-00155},
pages = {63 - 71},
year = {2019},
abstract = {CRISPR/Cas9-mediated gene editing of stem cells and primary
cell types has several limitations for clinical
applications. The direct delivery of ribonucleoprotein (RNP)
complexes consisting of Cas9 nuclease and guide RNA (gRNA)
has improved DNA- and virus-free gene modifications, but it
does not enable the essential enrichment of the gene-edited
cells. Here, we established a protocol for the fluorescent
labeling and delivery of CRISPR/Cas9-gRNA RNP in primary
human hematopoietic stem and progenitor cells (HSPCs) and
induced pluripotent stem cells (iPSCs). As a proof of
principle for genes with low-abundance transcripts and
context-dependent inducible expression, we successfully
deleted growth arrest and DNA-damage-inducible β (GADD45B).
We found that GADD45B is indispensable for DNA damage
protection and survival in stem cells. Thus, we describe an
easy and efficient protocol of DNA-free gene editing of
hard-to-target transcripts and enrichment of gene-modified
cells that are generally difficult to transfect.},
keywords = {Antigens, Differentiation (NLM Chemicals) / GADD45B
protein, human (NLM Chemicals) / Macromolecular Substances
(NLM Chemicals) / RNA, Guide (NLM Chemicals) /
Ribonucleoproteins (NLM Chemicals)},
cin = {L801},
ddc = {610},
cid = {I:(DE-He78)L801-20160331},
pnm = {899 - ohne Topic (POF3-899)},
pid = {G:(DE-HGF)POF3-899},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:30622144},
pmc = {pmc:PMC6325296},
doi = {10.1182/bloodadvances.2017015511},
url = {https://inrepo02.dkfz.de/record/153073},
}