Protocol for in vivo CRISPR knockout screening of autophagy genes to identify breast cancer metastasis suppressors
Summary: Metastasis represents the primary cause of mortality among patients with breast cancer. Here, we present a protocol for utilizing an in vivo custom CRISPR-Cas9 knockout library to systematically investigate autophagy regulatory genes implicated in breast cancer metastasis to the lung in mic...
Saved in:
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-06-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725002047 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849324767592579072 |
|---|---|
| author | Mingang Hao Peixin Lu Jun-Lin Guan |
| author_facet | Mingang Hao Peixin Lu Jun-Lin Guan |
| author_sort | Mingang Hao |
| collection | DOAJ |
| description | Summary: Metastasis represents the primary cause of mortality among patients with breast cancer. Here, we present a protocol for utilizing an in vivo custom CRISPR-Cas9 knockout library to systematically investigate autophagy regulatory genes implicated in breast cancer metastasis to the lung in mice. We describe steps for library synthesis, cloning, and virus packaging and transfection. We then detail procedures for genome DNA collection and transplantation, followed by analysis of screening data. This protocol enables efficient identification of potential suppressors of breast cancer metastasis in vivo.For complete details on the use and execution of this protocol, please refer to Hao et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-1deeee03989b4c4d8a27e481b93c62fc |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-1deeee03989b4c4d8a27e481b93c62fc2025-08-20T03:48:36ZengElsevierSTAR Protocols2666-16672025-06-016210379810.1016/j.xpro.2025.103798Protocol for in vivo CRISPR knockout screening of autophagy genes to identify breast cancer metastasis suppressorsMingang Hao0Peixin Lu1Jun-Lin Guan2Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA; Corresponding authorDepartment of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USADepartment of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USASummary: Metastasis represents the primary cause of mortality among patients with breast cancer. Here, we present a protocol for utilizing an in vivo custom CRISPR-Cas9 knockout library to systematically investigate autophagy regulatory genes implicated in breast cancer metastasis to the lung in mice. We describe steps for library synthesis, cloning, and virus packaging and transfection. We then detail procedures for genome DNA collection and transplantation, followed by analysis of screening data. This protocol enables efficient identification of potential suppressors of breast cancer metastasis in vivo.For complete details on the use and execution of this protocol, please refer to Hao et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725002047Cell BiologyCancerCRISPR |
| spellingShingle | Mingang Hao Peixin Lu Jun-Lin Guan Protocol for in vivo CRISPR knockout screening of autophagy genes to identify breast cancer metastasis suppressors STAR Protocols Cell Biology Cancer CRISPR |
| title | Protocol for in vivo CRISPR knockout screening of autophagy genes to identify breast cancer metastasis suppressors |
| title_full | Protocol for in vivo CRISPR knockout screening of autophagy genes to identify breast cancer metastasis suppressors |
| title_fullStr | Protocol for in vivo CRISPR knockout screening of autophagy genes to identify breast cancer metastasis suppressors |
| title_full_unstemmed | Protocol for in vivo CRISPR knockout screening of autophagy genes to identify breast cancer metastasis suppressors |
| title_short | Protocol for in vivo CRISPR knockout screening of autophagy genes to identify breast cancer metastasis suppressors |
| title_sort | protocol for in vivo crispr knockout screening of autophagy genes to identify breast cancer metastasis suppressors |
| topic | Cell Biology Cancer CRISPR |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725002047 |
| work_keys_str_mv | AT minganghao protocolforinvivocrisprknockoutscreeningofautophagygenestoidentifybreastcancermetastasissuppressors AT peixinlu protocolforinvivocrisprknockoutscreeningofautophagygenestoidentifybreastcancermetastasissuppressors AT junlinguan protocolforinvivocrisprknockoutscreeningofautophagygenestoidentifybreastcancermetastasissuppressors |