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The outbreak of the novel coronavirus pneumonia (COVID-19) has triggered a serious public health crisis worldwide
.
SARS-CoV-2 is the main pathogen that causes COVID-19.
At present, many studies have found key host factors related to SARS-CoV-2 infection, such as high-throughput functional screening based on the CRISPR gene knockout system
.
However, none of these screenings found new receptors other than ACE2, which may be because such loss-of-function-based screenings were performed in cell types where the expression and function of ACE2 were dominant [2-5]
On August 20, 2021, Peking University Wei Wensheng's group, Chinese Academy of Medical Sciences/Peking Union Medical College Wang Jianwei's group, and Peking University Xiao Junyu's group jointly published a titled Genome-wide CRISPR activation screen identifies candidate receptors in Science China Life Sciences.
For SARS-CoV-2 entry research paper, this study found a number of potential new cells that mediate SARS-CoV-2 invasion through genome-wide gain-of-function screening based on the CRISPR activation (CRISPRa) system Receptor
.
In order to systematically study the key factors of SARS-CoV-2 cell entry, the study used SARS-CoV-2 pseudovirus to perform genome-wide CRISPRa screening in HEK293T cells
.
The screening is based on the built-in molecular bar code (iBAR) method established in the laboratory [6] adding an external bar code (eBAR) to sgRNA to build a sgRNA eBAR library, so that high-infection multiplicity and a small amount of cells can be used for high-quality libraries Build
Figure 1 Through genome-wide CRISPRa functional screening, key host factors that affect SARS-CoV-2's entry into the cell were discovered
Combining the SARS-CoV-2 pseudovirus and true virus experiments, the study found that the membrane proteins LDLRAD3, TMEM30A and CLEC4G can effectively mediate virus invasion into cells in a way that is independent of ACE2
.
In addition, studies have confirmed that these membrane proteins can bind to SARS-CoV-2 S protein
Figure 2 SARS-CoV-2 infection experiments confirmed that LDLRAD3, CLEC4G and TMEM30A are potential receptors for mediating SARS-CoV-2 into the cell
Dr.
Zhu Shiyou, Dr.
Liu Ying, associate researcher Dr.
Zhou Zhuo and doctoral student Zhang Zhiying of Xiao Junyu's research group are the co-first authors of the paper
.
The research project has won the National Key Research and Development Program, the National Natural Science Foundation of China, the National Science and Technology Major Project for the Prevention and Control of Infectious Diseases, the Beijing Municipal Science and Technology Commission's Life Science Frontier Innovation and Cultivation Project, the Beijing Future Gene Diagnosis Advanced Innovation Center, and the Peking University-Tsinghua Life Science Joint Support from the center and the China Postdoctoral Science Foundation
Original link: http://engine.
references:
1.
2.
3.
4.
5.
6.
Zhu, S.
, Cao, Z.
, Liu, Z.
, He, Y.
, Wang, Y.
, Yuan, P.
, Li, W.
, Tian, F.
, Bao, Y.
, and Wei, W.
(2019).
Guide RNAs with embedded barcodes boost CRISPR-pooled screens.
Genome Biol 20 , 20.
