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On-line Access: 2024-08-27
Received: 2023-10-17
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Can SpRY recognize any PAM in human cells?
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Jinbin YE, Haitao XI, Yilu CHEN, Qishu CHEN, Xiaosheng LU, Jineng LV, Yamin CHEN, Feng GU, Junzhao ZHAO. Can SpRY recognize any PAM in human cells?[J]. Journal of Zhejiang University Science B, 2022, 23(5): 382-391.
@article{title="Can SpRY recognize any PAM in human cells?",
author="Jinbin YE, Haitao XI, Yilu CHEN, Qishu CHEN, Xiaosheng LU, Jineng LV, Yamin CHEN, Feng GU, Junzhao ZHAO",
journal="Journal of Zhejiang University Science B",
volume="23",
number="5",
pages="382-391",
year="2022",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B2100710"
}
%0 Journal Article
%T Can SpRY recognize any PAM in human cells?
%A Jinbin YE
%A Haitao XI
%A Yilu CHEN
%A Qishu CHEN
%A Xiaosheng LU
%A Jineng LV
%A Yamin CHEN
%A Feng GU
%A Junzhao ZHAO
%J Journal of Zhejiang University SCIENCE B
%V 23
%N 5
%P 382-391
%@ 1673-1581
%D 2022
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B2100710
TY - JOUR
T1 - Can SpRY recognize any PAM in human cells?
A1 - Jinbin YE
A1 - Haitao XI
A1 - Yilu CHEN
A1 - Qishu CHEN
A1 - Xiaosheng LU
A1 - Jineng LV
A1 - Yamin CHEN
A1 - Feng GU
A1 - Junzhao ZHAO
J0 - Journal of Zhejiang University Science B
VL - 23
IS - 5
SP - 382
EP - 391
%@ 1673-1581
Y1 - 2022
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B2100710
Abstract: The application of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) can be limited due to a lack of compatible protospacer adjacent motif (PAM) sequences in the DNA regions of interest. Recently, spRY, a variant of Streptococcus pyogenes Cas9 (SpCas9), was reported, which nearly completely fulfils the PAM requirement. Meanwhile, PAMs for spRY have not been well addressed. In our previous study, we developed the PAM Definition by Observable Sequence Excision (PAM-DOSE) and green fluorescent protein (GFP)-reporter systems to study PAMs in human cells. Herein, we endeavored to identify the PAMs of spRY with these two methods. The results indicated that 5'-NRN-3', 5'-NTA-3', and 5'-NCK-3' could be considered as canonical PAMs. 5'-NCA-3' and 5'-NTK-3' may serve as non-priority PAMs. At the same time, PAM of 5'-NYC-3' is not recommended for human cells. These findings provide further insights into the application of spRY for human genome editing.
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