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Updated: Oct 23, 2025
Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens
Published on: June 6, 2020
Genome-wide CRISPR activation screen identifies candidate receptors for SARS-CoV-2 entry
Shiyou Zhu1, Ying Liu1, Zhuo Zhou1
1Biomedical Pioneering Innovation Center, Beijing Advanced Innovation Center for Genomics, Peking-Tsinghua Center for Life Sciences, Peking University Genome Editing Research Center, State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, 100871, China.
Abstract:
The outbreak of coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 has created a global health crisis. SARS-CoV-2 infects varieties of tissues where the known receptor ACE2 is low or almost absent, suggesting the existence of alternative viral entry pathways. Here, we performed a genome-wide barcoded-CRISPRa screen to identify novel host factors that enable SARS-CoV-2 infection. Beyond known host proteins, i.e., ACE2, TMPRSS2, and NRP1, we identified multiple host components, among which LDLRAD3, TMEM30A, and CLEC4G were confirmed as functional receptors for SARS-CoV-2. All these membrane proteins bind directly to spike's N-terminal domain (NTD). Their essential and physiological roles have been confirmed in either neuron or liver cells. In particular, LDLRAD3 and CLEC4G mediate SARS-CoV-2 entry and infection in an ACE2-independent fashion. The identification of the novel receptors and entry mechanisms could advance our understanding of the multiorgan tropism of SARS-CoV-2, and may shed light on the development of COVID-19 countermeasures.
Insights
Researchers discovered new ways SARS-CoV-2 enters cells, identifying novel receptors like LDLRAD3 and CLEC4G that work independently of ACE2. This finding advances understanding of COVID-19
Area of Science:
- Virology
- Molecular Biology
- Cell Biology
Background:
- The COVID-19 pandemic, caused by SARS-CoV-2, highlights the need to understand viral entry mechanisms.
- SARS-CoV-2 infects tissues with low ACE2 expression, suggesting alternative entry pathways exist.
Purpose of the Study:
- To identify novel host factors and receptors facilitating SARS-CoV-2 infection.
- To elucidate ACE2-independent viral entry routes.
Main Methods:
- Genome-wide barcoded-CRISPRa screen to identify host factors.
- Confirmation of functional receptors through binding assays and infection studies.
- Investigation of viral entry in neuron and liver cells.
Main Results:
- Identified LDLRAD3, TMEM30A, and CLEC4G as functional SARS-CoV-2 receptors.
- These receptors bind directly to the spike protein's N-terminal domain (NTD).
- LDLRAD3 and CLEC4G mediate ACE2-independent SARS-CoV-2 entry and infection.
Conclusions:
- Novel receptors and entry mechanisms contribute to SARS-CoV-2's multiorgan tropism.
- Understanding these pathways is crucial for developing effective COVID-19 countermeasures.
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