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김소정
유전체 교정 연구단
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CRISPR RNAs trigger innate immune responses in human cells Highly Cited Paper

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Title
CRISPR RNAs trigger innate immune responses in human cells
Author(s)
Sojung Kim; Taeyoung Koo; Hyeon-Gun Jee; Hee-Yeon Cho; Gyeorae Lee; Dong-Gyun Lim; Hyoung Shik Shin; Jin-Soo Kim
Publication Date
2018-03
Journal
GENOME RESEARCH, v.28, no.3, pp.367 - 373
Publisher
Cold Spring Harbor Laboratory Press
Abstract
Here, we report that CRISPR guide RNAs (gRNAs) with a 5′-triphosphate group (5′-ppp gRNAs) produced via in vitro transcription trigger RNA-sensing innate immune responses in human and murine cells, leading to cytotoxicity. 5′-ppp gRNAs in the cytosol are recognized by DDX58, which in turn activates type I interferon responses, causing up to ∼80% cell death. We show that the triphosphate group can be removed by a phosphatase in vitro and that the resulting 5′-hydroxyl gRNAs in complex with Cas9 or Cpf1 avoid innate immune responses and can achieve targeted mutagenesis at a frequency of 95% in primary human CD4+ T cells. These results are in line with previous findings that chemically synthesized sgRNAs with a 5′-hydroxyl group are much more efficient than in vitro-transcribed (IVT) sgRNAs in human and other mammalian cells. The phosphatase treatment of IVT sgRNAs is a cost-effective method for making highly active sgRNAs, avoiding innate immune responses in human cells. © 2018 Kim et al
URI
https://pr.ibs.re.kr/handle/8788114/4601
ISSN
1088-9051
Appears in Collections:
Center for Genome Engineering(유전체 교정 연구단) > Journal Papers (저널논문)
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