CRISPR/Cas-based customization of pooled CRISPR libraries
Pooled CRISPR libraries are widely used in high-throughput screening to study various biological processes. Various pooled CRISPR libraries have been shared for CRISPR screens and useful tools have been developed to construct researcher's own libraries, however, many researchers are struggling...
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| Veröffentlicht in: | PloS one 2018-06, Vol.13 (6), p.e0199473-e0199473 |
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| creator | Kweon, Jiyeon Kim, Da-Eun Jang, An-Hee Kim, Yongsub |
| description | Pooled CRISPR libraries are widely used in high-throughput screening to study various biological processes. Various pooled CRISPR libraries have been shared for CRISPR screens and useful tools have been developed to construct researcher's own libraries, however, many researchers are struggling to create their own pooled CRISPR libraries: it is a time-consuming, labor-intensive, and expensive process. In this study, we develop a simple method to customize conventional pooled CRISPR libraries using the CRISPR/Cas9 system. We show that conventional pooled CRISPR libraries can be modified by eliminating gRNAs that target positive genes, enabling the identification of unknown target genes in CRISPR screening. CRISPR/Cas9 system can be applied as a precise tool for customizing conventional pooled CRISPR libraries and will broaden the scope of high-throughput screening technology. |
| doi_str_mv | 10.1371/journal.pone.0199473 |
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Various pooled CRISPR libraries have been shared for CRISPR screens and useful tools have been developed to construct researcher's own libraries, however, many researchers are struggling to create their own pooled CRISPR libraries: it is a time-consuming, labor-intensive, and expensive process. In this study, we develop a simple method to customize conventional pooled CRISPR libraries using the CRISPR/Cas9 system. We show that conventional pooled CRISPR libraries can be modified by eliminating gRNAs that target positive genes, enabling the identification of unknown target genes in CRISPR screening. CRISPR/Cas9 system can be applied as a precise tool for customizing conventional pooled CRISPR libraries and will broaden the scope of high-throughput screening technology.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0199473</identifier><identifier>PMID: 29924860</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Base Sequence ; Bioengineering ; Biological activity ; Biology and Life Sciences ; CRISPR ; CRISPR-Associated Protein 9 - metabolism ; CRISPR-Cas Systems - genetics ; Customization ; Deoxyribonucleic acid ; DNA ; Drug resistance ; Engineering and Technology ; FDA approval ; Gene Library ; Genes ; Genetic engineering ; Genomes ; Genomics ; HEK293 Cells ; HeLa Cells ; High-throughput screening ; High-Throughput Screening Assays ; Humans ; Hypoxanthine Phosphoribosyltransferase - metabolism ; Kinases ; Libraries ; Medical research ; Medicine ; Plasmids ; Proteins ; Research and Analysis Methods ; Ribonucleoproteins - metabolism ; Screening ; Stem cells ; Target recognition ; Technology application</subject><ispartof>PloS one, 2018-06, Vol.13 (6), p.e0199473-e0199473</ispartof><rights>COPYRIGHT 2018 Public Library of Science</rights><rights>2018 Kweon et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2018 Kweon et al 2018 Kweon et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-2fb003e244b0077befd57cbc41de0284497752cfc072147bb21ffe3610e3493e3</citedby><cites>FETCH-LOGICAL-c692t-2fb003e244b0077befd57cbc41de0284497752cfc072147bb21ffe3610e3493e3</cites><orcidid>0000-0002-9334-4333</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6010251/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6010251/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,725,778,782,862,883,2098,2917,23849,27907,27908,53774,53776,79351,79352</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29924860$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Maas, Stefan</contributor><creatorcontrib>Kweon, Jiyeon</creatorcontrib><creatorcontrib>Kim, Da-Eun</creatorcontrib><creatorcontrib>Jang, An-Hee</creatorcontrib><creatorcontrib>Kim, Yongsub</creatorcontrib><title>CRISPR/Cas-based customization of pooled CRISPR libraries</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Pooled CRISPR libraries are widely used in high-throughput screening to study various biological processes. Various pooled CRISPR libraries have been shared for CRISPR screens and useful tools have been developed to construct researcher's own libraries, however, many researchers are struggling to create their own pooled CRISPR libraries: it is a time-consuming, labor-intensive, and expensive process. In this study, we develop a simple method to customize conventional pooled CRISPR libraries using the CRISPR/Cas9 system. We show that conventional pooled CRISPR libraries can be modified by eliminating gRNAs that target positive genes, enabling the identification of unknown target genes in CRISPR screening. CRISPR/Cas9 system can be applied as a precise tool for customizing conventional pooled CRISPR libraries and will broaden the scope of high-throughput screening technology.</description><subject>Base Sequence</subject><subject>Bioengineering</subject><subject>Biological activity</subject><subject>Biology and Life Sciences</subject><subject>CRISPR</subject><subject>CRISPR-Associated Protein 9 - metabolism</subject><subject>CRISPR-Cas Systems - genetics</subject><subject>Customization</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>Drug resistance</subject><subject>Engineering and Technology</subject><subject>FDA approval</subject><subject>Gene Library</subject><subject>Genes</subject><subject>Genetic engineering</subject><subject>Genomes</subject><subject>Genomics</subject><subject>HEK293 Cells</subject><subject>HeLa Cells</subject><subject>High-throughput screening</subject><subject>High-Throughput Screening Assays</subject><subject>Humans</subject><subject>Hypoxanthine Phosphoribosyltransferase - metabolism</subject><subject>Kinases</subject><subject>Libraries</subject><subject>Medical research</subject><subject>Medicine</subject><subject>Plasmids</subject><subject>Proteins</subject><subject>Research and Analysis Methods</subject><subject>Ribonucleoproteins - metabolism</subject><subject>Screening</subject><subject>Stem cells</subject><subject>Target recognition</subject><subject>Technology application</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNqNkl2L1DAUhoso7rr6D0QHBNGLmT35aNPcCMvgx8DCyqx6G9L0dCZLphmTVtRfb-p0l6nsheQi4eQ578k5ebPsOYEFYYKc3_g-tNot9r7FBRApuWAPslMiGZ0XFNjDo_NJ9iTGG4CclUXxODuhUlJeFnCayeV6df15fb7UcV7piPXM9LHzO_tbd9a3M9_M9t67FD-AM2eroIPF-DR71GgX8dm4n2VfP7z_svw0v7z6uFpeXM5NIWk3p00FwJBynnYhKmzqXJjKcFIj0JJzKUROTWNAUMJFVVHSNMgKAsi4ZMjOspcH3b3zUY1dR0WhoEJSSSARqwNRe32j9sHudPilvLbqb8CHjdKhs8ahqqualFSkcZXIqSxkoYWsQeaSlSBlnrTejdX6aoe1wbYL2k1Epzet3aqN_6EKIEBzkgTejALBf-8xdmpno0HndIu-H96di1IQIEOtV_-g93c3UhudGrBt41NdM4iqi5wzQQGAJmpxD5VWjTtrkkUam-KThLeThMR0-LPb6D5Gtbpe_z979W3Kvj5it6hdt43e9YOb4hTkB9AEH2PA5m7IBNTg8NtpqMHhanR4Sntx_EF3SbeWZn8AqvPxqw</recordid><startdate>20180620</startdate><enddate>20180620</enddate><creator>Kweon, Jiyeon</creator><creator>Kim, Da-Eun</creator><creator>Jang, An-Hee</creator><creator>Kim, Yongsub</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</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>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0002-9334-4333</orcidid></search><sort><creationdate>20180620</creationdate><title>CRISPR/Cas-based customization of pooled CRISPR libraries</title><author>Kweon, Jiyeon ; Kim, Da-Eun ; Jang, An-Hee ; Kim, Yongsub</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-2fb003e244b0077befd57cbc41de0284497752cfc072147bb21ffe3610e3493e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Base Sequence</topic><topic>Bioengineering</topic><topic>Biological activity</topic><topic>Biology and Life Sciences</topic><topic>CRISPR</topic><topic>CRISPR-Associated Protein 9 - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kweon, Jiyeon</au><au>Kim, Da-Eun</au><au>Jang, An-Hee</au><au>Kim, Yongsub</au><au>Maas, Stefan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>CRISPR/Cas-based customization of pooled CRISPR libraries</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2018-06-20</date><risdate>2018</risdate><volume>13</volume><issue>6</issue><spage>e0199473</spage><epage>e0199473</epage><pages>e0199473-e0199473</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Pooled CRISPR libraries are widely used in high-throughput screening to study various biological processes. Various pooled CRISPR libraries have been shared for CRISPR screens and useful tools have been developed to construct researcher's own libraries, however, many researchers are struggling to create their own pooled CRISPR libraries: it is a time-consuming, labor-intensive, and expensive process. In this study, we develop a simple method to customize conventional pooled CRISPR libraries using the CRISPR/Cas9 system. We show that conventional pooled CRISPR libraries can be modified by eliminating gRNAs that target positive genes, enabling the identification of unknown target genes in CRISPR screening. CRISPR/Cas9 system can be applied as a precise tool for customizing conventional pooled CRISPR libraries and will broaden the scope of high-throughput screening technology.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>29924860</pmid><doi>10.1371/journal.pone.0199473</doi><tpages>e0199473</tpages><orcidid>https://orcid.org/0000-0002-9334-4333</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Base Sequence Bioengineering Biological activity Biology and Life Sciences CRISPR CRISPR-Associated Protein 9 - metabolism CRISPR-Cas Systems - genetics Customization Deoxyribonucleic acid DNA Drug resistance Engineering and Technology FDA approval Gene Library Genes Genetic engineering Genomes Genomics HEK293 Cells HeLa Cells High-throughput screening High-Throughput Screening Assays Humans Hypoxanthine Phosphoribosyltransferase - metabolism Kinases Libraries Medical research Medicine Plasmids Proteins Research and Analysis Methods Ribonucleoproteins - metabolism Screening Stem cells Target recognition Technology application |
| title | CRISPR/Cas-based customization of pooled CRISPR libraries |
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