Identifying genome-wide off-target sites of CRISPR RNA–guided nucleases and deaminases with Digenome-seq
Digested genome sequencing (Digenome-seq) is a highly sensitive, easy-to-carry-out, cell-free method for experimentally identifying genome-wide off-target sites of programmable nucleases and deaminases (also known as base editors). Genomic DNA is digested in vitro using clustered regularly interspac...
Gespeichert in:
| Veröffentlicht in: | Nature protocols 2021-02, Vol.16 (2), p.1170-1192 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 1192 |
|---|---|
| container_issue | 2 |
| container_start_page | 1170 |
| container_title | Nature protocols |
| container_volume | 16 |
| creator | Kim, Daesik Kang, Beum-Chang Kim, Jin-Soo |
| description | Digested genome sequencing (Digenome-seq) is a highly sensitive, easy-to-carry-out, cell-free method for experimentally identifying genome-wide off-target sites of programmable nucleases and deaminases (also known as base editors). Genomic DNA is digested in vitro using clustered regularly interspaced short palindromic repeats ribonucleoproteins (RNPs; plus DNA-modifying enzymes to cleave both strands of DNA at sites containing deaminated base products, in the case of base editors) and subjected to whole-genome sequencing (WGS) with a typical sequencing depth of 30×. A web-based program is available to map in vitro cleavage sites corresponding to on- and off-target sites. Chromatin DNA, in parallel with histone-free genomic DNA, can also be used to account for the effects of chromatin structure on off-target nuclease activity. Digenome-seq is more sensitive and comprehensive than cell-based methods for identifying off-target sites. Unlike other cell-free methods, Digenome-seq does not involve enrichment of DNA ends through PCR amplification. The entire process other than WGS, which takes ~1–2 weeks, including purification and preparation of RNPs, digestion of genomic DNA and bioinformatic analysis after WGS, takes about several weeks.
This protocol describes a cell-free method for experimentally identifying genome-wide off-target sites of CRISPR nucleases and deaminases through in vitro digestion of genomic DNA or chromatin followed by whole-genome sequencing. |
| doi_str_mv | 10.1038/s41596-020-00453-6 |
| format | Article |
| fullrecord | <record><control><sourceid>gale_proqu</sourceid><recordid>TN_cdi_proquest_miscellaneous_2479040579</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A655717208</galeid><sourcerecordid>A655717208</sourcerecordid><originalsourceid>FETCH-LOGICAL-c476t-72315358a8dc4d76c3d71c19c7672664b8c00e16a64d8583ac567e62751050ad3</originalsourceid><addsrcrecordid>eNp9ks1u1DAUhSMEoqXwAixQJDawcPG_k-Vo-BupAjSFteXaN8GjxGltR6U73oE35ElwZwaqQQh5Yfv6O0e-9qmqpwSfEsyaV4kT0UqEKUYYc8GQvFcdEyUwoqpt72_XHFHStEfVo5Q2BVJMqofVEWNcUs7a42qzchCy72586OsewjQCuvYO6qnrUDaxh1wnnyGVQr1cr84_rev1h8XP7z_6uWCuDrMdwKQCmOBqB2b0Ybu99vlr_drvPRNcPa4edGZI8GQ_n1Rf3r75vHyPzj6-Wy0XZ8hyJTNSlBHBRGMaZ7lT0jKniCWtVVJRKflFYzEGIo3krhENM1ZIBZIqQbDAxrGT6sXO9zJOVzOkrEefLAyDCTDNSVOuWsyxUG1Bn_-FbqY5hnK7QjWKSFFe6Y7qzQDah27K0dhbU72QQiiiKG4KdfoPqgwHo7dTgM6X-oHg5YGgMBm-5d7MKenV-fqQpTvWximlCJ2-jH408UYTrG_DoHdh0CUMehsGLYvo2b67-WIE90fy-_cLwHZAKkehh3jX_n9sfwGI2bvu</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2487165243</pqid></control><display><type>article</type><title>Identifying genome-wide off-target sites of CRISPR RNA–guided nucleases and deaminases with Digenome-seq</title><source>MEDLINE</source><source>Springer Nature - Complete Springer Journals</source><source>Nature Journals Online</source><creator>Kim, Daesik ; Kang, Beum-Chang ; Kim, Jin-Soo</creator><creatorcontrib>Kim, Daesik ; Kang, Beum-Chang ; Kim, Jin-Soo</creatorcontrib><description>Digested genome sequencing (Digenome-seq) is a highly sensitive, easy-to-carry-out, cell-free method for experimentally identifying genome-wide off-target sites of programmable nucleases and deaminases (also known as base editors). Genomic DNA is digested in vitro using clustered regularly interspaced short palindromic repeats ribonucleoproteins (RNPs; plus DNA-modifying enzymes to cleave both strands of DNA at sites containing deaminated base products, in the case of base editors) and subjected to whole-genome sequencing (WGS) with a typical sequencing depth of 30×. A web-based program is available to map in vitro cleavage sites corresponding to on- and off-target sites. Chromatin DNA, in parallel with histone-free genomic DNA, can also be used to account for the effects of chromatin structure on off-target nuclease activity. Digenome-seq is more sensitive and comprehensive than cell-based methods for identifying off-target sites. Unlike other cell-free methods, Digenome-seq does not involve enrichment of DNA ends through PCR amplification. The entire process other than WGS, which takes ~1–2 weeks, including purification and preparation of RNPs, digestion of genomic DNA and bioinformatic analysis after WGS, takes about several weeks.
This protocol describes a cell-free method for experimentally identifying genome-wide off-target sites of CRISPR nucleases and deaminases through in vitro digestion of genomic DNA or chromatin followed by whole-genome sequencing.</description><identifier>ISSN: 1754-2189</identifier><identifier>EISSN: 1750-2799</identifier><identifier>DOI: 10.1038/s41596-020-00453-6</identifier><identifier>PMID: 33462439</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>631/1647/514/2254 ; 631/208/4041/3196 ; 631/61/514/1948 ; Analytical Chemistry ; Base Sequence ; Biological Techniques ; Biomedical and Life Sciences ; Chromatin ; Chromosome Mapping - methods ; Clustered Regularly Interspaced Short Palindromic Repeats ; Computational Biology/Bioinformatics ; CRISPR ; CRISPR-Associated Protein 9 - metabolism ; CRISPR-Cas Systems ; Deoxyribonucleic acid ; Digestion ; DNA ; DNA sequencing ; Endonucleases - metabolism ; Gene Editing - methods ; Genome, Human ; Genomes ; Genomics ; High-Throughput Nucleotide Sequencing - methods ; Histones ; Humans ; Identification and classification ; Kinases ; Life Sciences ; Methods ; Microarrays ; Nuclease ; Nucleases ; Nucleotide Deaminases - genetics ; Nucleotide Deaminases - metabolism ; Nucleotide sequencing ; Organic Chemistry ; Protocol ; Ribonucleases - genetics ; Ribonucleases - metabolism ; Ribonucleic acid ; Ribonucleoproteins ; RNA ; RNA, Guide, CRISPR-Cas Systems - genetics ; Sequence Analysis, DNA - methods ; Structure ; Whole genome sequencing ; Whole Genome Sequencing - methods</subject><ispartof>Nature protocols, 2021-02, Vol.16 (2), p.1170-1192</ispartof><rights>The Author(s), under exclusive licence to Springer Nature Limited 2021</rights><rights>COPYRIGHT 2021 Nature Publishing Group</rights><rights>The Author(s), under exclusive licence to Springer Nature Limited 2021.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c476t-72315358a8dc4d76c3d71c19c7672664b8c00e16a64d8583ac567e62751050ad3</citedby><cites>FETCH-LOGICAL-c476t-72315358a8dc4d76c3d71c19c7672664b8c00e16a64d8583ac567e62751050ad3</cites><orcidid>0000-0003-4847-1306 ; 0000-0003-0806-5979</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/s41596-020-00453-6$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/s41596-020-00453-6$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33462439$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kim, Daesik</creatorcontrib><creatorcontrib>Kang, Beum-Chang</creatorcontrib><creatorcontrib>Kim, Jin-Soo</creatorcontrib><title>Identifying genome-wide off-target sites of CRISPR RNA–guided nucleases and deaminases with Digenome-seq</title><title>Nature protocols</title><addtitle>Nat Protoc</addtitle><addtitle>Nat Protoc</addtitle><description>Digested genome sequencing (Digenome-seq) is a highly sensitive, easy-to-carry-out, cell-free method for experimentally identifying genome-wide off-target sites of programmable nucleases and deaminases (also known as base editors). Genomic DNA is digested in vitro using clustered regularly interspaced short palindromic repeats ribonucleoproteins (RNPs; plus DNA-modifying enzymes to cleave both strands of DNA at sites containing deaminated base products, in the case of base editors) and subjected to whole-genome sequencing (WGS) with a typical sequencing depth of 30×. A web-based program is available to map in vitro cleavage sites corresponding to on- and off-target sites. Chromatin DNA, in parallel with histone-free genomic DNA, can also be used to account for the effects of chromatin structure on off-target nuclease activity. Digenome-seq is more sensitive and comprehensive than cell-based methods for identifying off-target sites. Unlike other cell-free methods, Digenome-seq does not involve enrichment of DNA ends through PCR amplification. The entire process other than WGS, which takes ~1–2 weeks, including purification and preparation of RNPs, digestion of genomic DNA and bioinformatic analysis after WGS, takes about several weeks.
This protocol describes a cell-free method for experimentally identifying genome-wide off-target sites of CRISPR nucleases and deaminases through in vitro digestion of genomic DNA or chromatin followed by whole-genome sequencing.</description><subject>631/1647/514/2254</subject><subject>631/208/4041/3196</subject><subject>631/61/514/1948</subject><subject>Analytical Chemistry</subject><subject>Base Sequence</subject><subject>Biological Techniques</subject><subject>Biomedical and Life Sciences</subject><subject>Chromatin</subject><subject>Chromosome Mapping - methods</subject><subject>Clustered Regularly Interspaced Short Palindromic Repeats</subject><subject>Computational Biology/Bioinformatics</subject><subject>CRISPR</subject><subject>CRISPR-Associated Protein 9 - metabolism</subject><subject>CRISPR-Cas Systems</subject><subject>Deoxyribonucleic acid</subject><subject>Digestion</subject><subject>DNA</subject><subject>DNA sequencing</subject><subject>Endonucleases - metabolism</subject><subject>Gene Editing - methods</subject><subject>Genome, Human</subject><subject>Genomes</subject><subject>Genomics</subject><subject>High-Throughput Nucleotide Sequencing - methods</subject><subject>Histones</subject><subject>Humans</subject><subject>Identification and classification</subject><subject>Kinases</subject><subject>Life Sciences</subject><subject>Methods</subject><subject>Microarrays</subject><subject>Nuclease</subject><subject>Nucleases</subject><subject>Nucleotide Deaminases - genetics</subject><subject>Nucleotide Deaminases - metabolism</subject><subject>Nucleotide sequencing</subject><subject>Organic Chemistry</subject><subject>Protocol</subject><subject>Ribonucleases - genetics</subject><subject>Ribonucleases - metabolism</subject><subject>Ribonucleic acid</subject><subject>Ribonucleoproteins</subject><subject>RNA</subject><subject>RNA, Guide, CRISPR-Cas Systems - genetics</subject><subject>Sequence Analysis, DNA - methods</subject><subject>Structure</subject><subject>Whole genome sequencing</subject><subject>Whole Genome Sequencing - methods</subject><issn>1754-2189</issn><issn>1750-2799</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNp9ks1u1DAUhSMEoqXwAixQJDawcPG_k-Vo-BupAjSFteXaN8GjxGltR6U73oE35ElwZwaqQQh5Yfv6O0e-9qmqpwSfEsyaV4kT0UqEKUYYc8GQvFcdEyUwoqpt72_XHFHStEfVo5Q2BVJMqofVEWNcUs7a42qzchCy72586OsewjQCuvYO6qnrUDaxh1wnnyGVQr1cr84_rev1h8XP7z_6uWCuDrMdwKQCmOBqB2b0Ybu99vlr_drvPRNcPa4edGZI8GQ_n1Rf3r75vHyPzj6-Wy0XZ8hyJTNSlBHBRGMaZ7lT0jKniCWtVVJRKflFYzEGIo3krhENM1ZIBZIqQbDAxrGT6sXO9zJOVzOkrEefLAyDCTDNSVOuWsyxUG1Bn_-FbqY5hnK7QjWKSFFe6Y7qzQDah27K0dhbU72QQiiiKG4KdfoPqgwHo7dTgM6X-oHg5YGgMBm-5d7MKenV-fqQpTvWximlCJ2-jH408UYTrG_DoHdh0CUMehsGLYvo2b67-WIE90fy-_cLwHZAKkehh3jX_n9sfwGI2bvu</recordid><startdate>20210201</startdate><enddate>20210201</enddate><creator>Kim, Daesik</creator><creator>Kang, Beum-Chang</creator><creator>Kim, Jin-Soo</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7T5</scope><scope>7T7</scope><scope>7TM</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PATMY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0003-4847-1306</orcidid><orcidid>https://orcid.org/0000-0003-0806-5979</orcidid></search><sort><creationdate>20210201</creationdate><title>Identifying genome-wide off-target sites of CRISPR RNA–guided nucleases and deaminases with Digenome-seq</title><author>Kim, Daesik ; Kang, Beum-Chang ; Kim, Jin-Soo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c476t-72315358a8dc4d76c3d71c19c7672664b8c00e16a64d8583ac567e62751050ad3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>631/1647/514/2254</topic><topic>631/208/4041/3196</topic><topic>631/61/514/1948</topic><topic>Analytical Chemistry</topic><topic>Base Sequence</topic><topic>Biological Techniques</topic><topic>Biomedical and Life Sciences</topic><topic>Chromatin</topic><topic>Chromosome Mapping - methods</topic><topic>Clustered Regularly Interspaced Short Palindromic Repeats</topic><topic>Computational Biology/Bioinformatics</topic><topic>CRISPR</topic><topic>CRISPR-Associated Protein 9 - metabolism</topic><topic>CRISPR-Cas Systems</topic><topic>Deoxyribonucleic acid</topic><topic>Digestion</topic><topic>DNA</topic><topic>DNA sequencing</topic><topic>Endonucleases - metabolism</topic><topic>Gene Editing - methods</topic><topic>Genome, Human</topic><topic>Genomes</topic><topic>Genomics</topic><topic>High-Throughput Nucleotide Sequencing - methods</topic><topic>Histones</topic><topic>Humans</topic><topic>Identification and classification</topic><topic>Kinases</topic><topic>Life Sciences</topic><topic>Methods</topic><topic>Microarrays</topic><topic>Nuclease</topic><topic>Nucleases</topic><topic>Nucleotide Deaminases - genetics</topic><topic>Nucleotide Deaminases - metabolism</topic><topic>Nucleotide sequencing</topic><topic>Organic Chemistry</topic><topic>Protocol</topic><topic>Ribonucleases - genetics</topic><topic>Ribonucleases - metabolism</topic><topic>Ribonucleic acid</topic><topic>Ribonucleoproteins</topic><topic>RNA</topic><topic>RNA, Guide, CRISPR-Cas Systems - genetics</topic><topic>Sequence Analysis, DNA - methods</topic><topic>Structure</topic><topic>Whole genome sequencing</topic><topic>Whole Genome Sequencing - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kim, Daesik</creatorcontrib><creatorcontrib>Kang, Beum-Chang</creatorcontrib><creatorcontrib>Kim, Jin-Soo</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Immunology Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Nature protocols</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kim, Daesik</au><au>Kang, Beum-Chang</au><au>Kim, Jin-Soo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identifying genome-wide off-target sites of CRISPR RNA–guided nucleases and deaminases with Digenome-seq</atitle><jtitle>Nature protocols</jtitle><stitle>Nat Protoc</stitle><addtitle>Nat Protoc</addtitle><date>2021-02-01</date><risdate>2021</risdate><volume>16</volume><issue>2</issue><spage>1170</spage><epage>1192</epage><pages>1170-1192</pages><issn>1754-2189</issn><eissn>1750-2799</eissn><abstract>Digested genome sequencing (Digenome-seq) is a highly sensitive, easy-to-carry-out, cell-free method for experimentally identifying genome-wide off-target sites of programmable nucleases and deaminases (also known as base editors). Genomic DNA is digested in vitro using clustered regularly interspaced short palindromic repeats ribonucleoproteins (RNPs; plus DNA-modifying enzymes to cleave both strands of DNA at sites containing deaminated base products, in the case of base editors) and subjected to whole-genome sequencing (WGS) with a typical sequencing depth of 30×. A web-based program is available to map in vitro cleavage sites corresponding to on- and off-target sites. Chromatin DNA, in parallel with histone-free genomic DNA, can also be used to account for the effects of chromatin structure on off-target nuclease activity. Digenome-seq is more sensitive and comprehensive than cell-based methods for identifying off-target sites. Unlike other cell-free methods, Digenome-seq does not involve enrichment of DNA ends through PCR amplification. The entire process other than WGS, which takes ~1–2 weeks, including purification and preparation of RNPs, digestion of genomic DNA and bioinformatic analysis after WGS, takes about several weeks.
This protocol describes a cell-free method for experimentally identifying genome-wide off-target sites of CRISPR nucleases and deaminases through in vitro digestion of genomic DNA or chromatin followed by whole-genome sequencing.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>33462439</pmid><doi>10.1038/s41596-020-00453-6</doi><tpages>23</tpages><orcidid>https://orcid.org/0000-0003-4847-1306</orcidid><orcidid>https://orcid.org/0000-0003-0806-5979</orcidid></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1754-2189 |
| ispartof | Nature protocols, 2021-02, Vol.16 (2), p.1170-1192 |
| issn | 1754-2189 1750-2799 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_2479040579 |
| source | MEDLINE; Springer Nature - Complete Springer Journals; Nature Journals Online |
| subjects | 631/1647/514/2254 631/208/4041/3196 631/61/514/1948 Analytical Chemistry Base Sequence Biological Techniques Biomedical and Life Sciences Chromatin Chromosome Mapping - methods Clustered Regularly Interspaced Short Palindromic Repeats Computational Biology/Bioinformatics CRISPR CRISPR-Associated Protein 9 - metabolism CRISPR-Cas Systems Deoxyribonucleic acid Digestion DNA DNA sequencing Endonucleases - metabolism Gene Editing - methods Genome, Human Genomes Genomics High-Throughput Nucleotide Sequencing - methods Histones Humans Identification and classification Kinases Life Sciences Methods Microarrays Nuclease Nucleases Nucleotide Deaminases - genetics Nucleotide Deaminases - metabolism Nucleotide sequencing Organic Chemistry Protocol Ribonucleases - genetics Ribonucleases - metabolism Ribonucleic acid Ribonucleoproteins RNA RNA, Guide, CRISPR-Cas Systems - genetics Sequence Analysis, DNA - methods Structure Whole genome sequencing Whole Genome Sequencing - methods |
| title | Identifying genome-wide off-target sites of CRISPR RNA–guided nucleases and deaminases with Digenome-seq |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-30T03%3A56%3A41IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_proqu&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Identifying%20genome-wide%20off-target%20sites%20of%20CRISPR%20RNA%E2%80%93guided%20nucleases%20and%20deaminases%20with%20Digenome-seq&rft.jtitle=Nature%20protocols&rft.au=Kim,%20Daesik&rft.date=2021-02-01&rft.volume=16&rft.issue=2&rft.spage=1170&rft.epage=1192&rft.pages=1170-1192&rft.issn=1754-2189&rft.eissn=1750-2799&rft_id=info:doi/10.1038/s41596-020-00453-6&rft_dat=%3Cgale_proqu%3EA655717208%3C/gale_proqu%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2487165243&rft_id=info:pmid/33462439&rft_galeid=A655717208&rfr_iscdi=true |