Protocol for high-throughput screening of functional lysine residues in cell fitness
Summary: Amino acid residues are crucial to protein structure and function and have links to various human diseases. Here, we present a protocol for screening functional lysine residues across the human genome. We describe steps for designing lysine codon-targeting single-guide RNAs (sgRNAs), constr...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-12-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724005835 |
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| author | Ying Bao Wensheng Wei |
| author_facet | Ying Bao Wensheng Wei |
| author_sort | Ying Bao |
| collection | DOAJ |
| description | Summary: Amino acid residues are crucial to protein structure and function and have links to various human diseases. Here, we present a protocol for screening functional lysine residues across the human genome. We describe steps for designing lysine codon-targeting single-guide RNAs (sgRNAs), constructing an sgRNA library, conducting cell fitness screenings, and acquiring screening results. This approach leverages base editing and high-throughput screening techniques to systematically examine functional amino acid residues.For complete details on the use and execution of this protocol, please refer to Bao 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-1e8d5ac7a8ea48aa9df9eedd9b4006b5 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-1e8d5ac7a8ea48aa9df9eedd9b4006b52024-11-20T05:08:08ZengElsevierSTAR Protocols2666-16672024-12-0154103418Protocol for high-throughput screening of functional lysine residues in cell fitnessYing Bao0Wensheng Wei1Changping Laboratory, Beijing 102206, ChinaChangping Laboratory, Beijing 102206, China; Biomedical Pioneering Innovation Center, 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; Corresponding authorSummary: Amino acid residues are crucial to protein structure and function and have links to various human diseases. Here, we present a protocol for screening functional lysine residues across the human genome. We describe steps for designing lysine codon-targeting single-guide RNAs (sgRNAs), constructing an sgRNA library, conducting cell fitness screenings, and acquiring screening results. This approach leverages base editing and high-throughput screening techniques to systematically examine functional amino acid residues.For complete details on the use and execution of this protocol, please refer to Bao 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/S2666166724005835high-throughput screeningmolecular biologyCRISPR |
| spellingShingle | Ying Bao Wensheng Wei Protocol for high-throughput screening of functional lysine residues in cell fitness STAR Protocols high-throughput screening molecular biology CRISPR |
| title | Protocol for high-throughput screening of functional lysine residues in cell fitness |
| title_full | Protocol for high-throughput screening of functional lysine residues in cell fitness |
| title_fullStr | Protocol for high-throughput screening of functional lysine residues in cell fitness |
| title_full_unstemmed | Protocol for high-throughput screening of functional lysine residues in cell fitness |
| title_short | Protocol for high-throughput screening of functional lysine residues in cell fitness |
| title_sort | protocol for high throughput screening of functional lysine residues in cell fitness |
| topic | high-throughput screening molecular biology CRISPR |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724005835 |
| work_keys_str_mv | AT yingbao protocolforhighthroughputscreeningoffunctionallysineresiduesincellfitness AT wenshengwei protocolforhighthroughputscreeningoffunctionallysineresiduesincellfitness |