In vivo genome editing using the Cpf1 ortholog derived from Eubacterium eligens
Cpf1 is an RNA-guided endonuclease that can be programmed to cleave DNA targets. Specific features, such as containing a short crRNA, creating a staggered cleavage pattern and having a low off-target rate, render Cpf1 a promising gene-editing tool. Here, we present a new Cpf1 ortholog, EeCpf1, as a...
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| Veröffentlicht in: | Scientific reports 2019-09, Vol.9 (1), p.13911-7, Article 13911 |
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| creator | Ahn, Woo-Chan Park, Kwang-Hyun Bak, In Seon Song, Hyung-Nam An, Yan Lee, Su-Jin Jung, Mira Yoo, Kyeong-Won Yu, Dae-Yeul Kim, Yong-Sam Oh, Byung-Ha Woo, Eui-Jeon |
| description | Cpf1 is an RNA-guided endonuclease that can be programmed to cleave DNA targets. Specific features, such as containing a short crRNA, creating a staggered cleavage pattern and having a low off-target rate, render Cpf1 a promising gene-editing tool. Here, we present a new Cpf1 ortholog, EeCpf1, as a genome-editing tool; this ortholog is derived from the gut bacterial species
Eubacterium eligens
. EeCpf1 exhibits a higher cleavage activity with the Mn
2+
metal cofactor and efficiently cuts the target DNA with an engineered, nucleotide extended crRNA at the 5′ target site. When mouse blastocysts were injected with multitargeting crRNAs against the IL2R-γ gene, an essential gene for immunodeficient mouse model production, EeCpf1 efficiently generated IL2R-γ knockout mice. For the first time, these results demonstrate that EeCpf1 can be used as an
in vivo
gene-editing tool for the production of knockout mice. The utilization of engineered crRNA with multiple target sites will help to explore the
in vivo
DNA cleavage activities of Cpf1 orthologs from other species that have not been demonstrated. |
| doi_str_mv | 10.1038/s41598-019-50423-6 |
| format | Article |
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Eubacterium eligens
. EeCpf1 exhibits a higher cleavage activity with the Mn
2+
metal cofactor and efficiently cuts the target DNA with an engineered, nucleotide extended crRNA at the 5′ target site. When mouse blastocysts were injected with multitargeting crRNAs against the IL2R-γ gene, an essential gene for immunodeficient mouse model production, EeCpf1 efficiently generated IL2R-γ knockout mice. For the first time, these results demonstrate that EeCpf1 can be used as an
in vivo
gene-editing tool for the production of knockout mice. The utilization of engineered crRNA with multiple target sites will help to explore the
in vivo
DNA cleavage activities of Cpf1 orthologs from other species that have not been demonstrated.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/s41598-019-50423-6</identifier><identifier>PMID: 31558757</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>38 ; 38/70 ; 42 ; 42/41 ; 631/1647/1511 ; 631/61/17/1511 ; Animals ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Blastocyst - metabolism ; Blastocysts ; Deoxyribonucleic acid ; DNA ; Endonuclease ; Endonucleases - genetics ; Endonucleases - metabolism ; Eubacterium - enzymology ; Gene Editing - methods ; Genome editing ; Genomes ; Humanities and Social Sciences ; Immunodeficiency ; Interleukin 2 receptors ; Interleukin Receptor Common gamma Subunit - genetics ; Interleukin Receptor Common gamma Subunit - metabolism ; Magnesium - metabolism ; Mice ; Mice, Inbred C57BL ; multidisciplinary ; Ribonucleic acid ; RNA ; RNA, Circular - genetics ; Rodents ; Science ; Science (multidisciplinary)</subject><ispartof>Scientific reports, 2019-09, Vol.9 (1), p.13911-7, Article 13911</ispartof><rights>The Author(s) 2019</rights><rights>2019. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c474t-9914117503be4bbb3a8772757f8e74dd4be8433fb1e3596f904ec1b8ece31bba3</citedby><cites>FETCH-LOGICAL-c474t-9914117503be4bbb3a8772757f8e74dd4be8433fb1e3596f904ec1b8ece31bba3</cites><orcidid>0000-0001-6983-0270 ; 0000-0002-6437-8470</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/PMC6763456/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6763456/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,27901,27902,41096,42165,51551,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31558757$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ahn, Woo-Chan</creatorcontrib><creatorcontrib>Park, Kwang-Hyun</creatorcontrib><creatorcontrib>Bak, In Seon</creatorcontrib><creatorcontrib>Song, Hyung-Nam</creatorcontrib><creatorcontrib>An, Yan</creatorcontrib><creatorcontrib>Lee, Su-Jin</creatorcontrib><creatorcontrib>Jung, Mira</creatorcontrib><creatorcontrib>Yoo, Kyeong-Won</creatorcontrib><creatorcontrib>Yu, Dae-Yeul</creatorcontrib><creatorcontrib>Kim, Yong-Sam</creatorcontrib><creatorcontrib>Oh, Byung-Ha</creatorcontrib><creatorcontrib>Woo, Eui-Jeon</creatorcontrib><title>In vivo genome editing using the Cpf1 ortholog derived from Eubacterium eligens</title><title>Scientific reports</title><addtitle>Sci Rep</addtitle><addtitle>Sci Rep</addtitle><description>Cpf1 is an RNA-guided endonuclease that can be programmed to cleave DNA targets. Specific features, such as containing a short crRNA, creating a staggered cleavage pattern and having a low off-target rate, render Cpf1 a promising gene-editing tool. Here, we present a new Cpf1 ortholog, EeCpf1, as a genome-editing tool; this ortholog is derived from the gut bacterial species
Eubacterium eligens
. EeCpf1 exhibits a higher cleavage activity with the Mn
2+
metal cofactor and efficiently cuts the target DNA with an engineered, nucleotide extended crRNA at the 5′ target site. When mouse blastocysts were injected with multitargeting crRNAs against the IL2R-γ gene, an essential gene for immunodeficient mouse model production, EeCpf1 efficiently generated IL2R-γ knockout mice. For the first time, these results demonstrate that EeCpf1 can be used as an
in vivo
gene-editing tool for the production of knockout mice. The utilization of engineered crRNA with multiple target sites will help to explore the
in vivo
DNA cleavage activities of Cpf1 orthologs from other species that have not been demonstrated.</description><subject>38</subject><subject>38/70</subject><subject>42</subject><subject>42/41</subject><subject>631/1647/1511</subject><subject>631/61/17/1511</subject><subject>Animals</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Blastocyst - metabolism</subject><subject>Blastocysts</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>Endonuclease</subject><subject>Endonucleases - genetics</subject><subject>Endonucleases - metabolism</subject><subject>Eubacterium - enzymology</subject><subject>Gene Editing - methods</subject><subject>Genome editing</subject><subject>Genomes</subject><subject>Humanities and Social Sciences</subject><subject>Immunodeficiency</subject><subject>Interleukin 2 receptors</subject><subject>Interleukin Receptor Common gamma Subunit - genetics</subject><subject>Interleukin Receptor Common gamma Subunit - metabolism</subject><subject>Magnesium - metabolism</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>multidisciplinary</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA, Circular - genetics</subject><subject>Rodents</subject><subject>Science</subject><subject>Science (multidisciplinary)</subject><issn>2045-2322</issn><issn>2045-2322</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNp9kU1L7DAUhoMoKuofcCEBN256zWebbAQZxnsFwY2uQ9OediJtMybtgP_e1PH6tTCLJOS858l7eBE6peQPJVxdRkGlVhmhOpNEMJ7lO-iQESEzxhnb_XI_QCcxPpG0JNOC6n10wKmUqpDFIbq_HfDGbTxuYfA9YKjd6IYWT3HexxXgxbqh2Idx5Tvf4hqC20CNm-B7vJxsWY3pZeoxdC4h4jHaa8ouwsn7eYQeb5YPi3_Z3f3f28X1XVaJQoyZ1lRQWkjCLQhrLS9VUbDkqFFQiLoWFpTgvLEUuNR5o4mAiloFFXBqbcmP0NWWu55sD3UFwxjKzqyD68vwYnzpzPfK4Fam9RuTFzkXMk-Ai3dA8M8TxNH0LlbQdeUAfoqGseSRayFpkp7_kD75KQxpvFmlqM6VmIFsq6qCjzFA82GGEjNHZraRmRSZeYvMzE1nX8f4aPkfUBLwrSCm0tBC-Pz7F-wrj06h4A</recordid><startdate>20190926</startdate><enddate>20190926</enddate><creator>Ahn, Woo-Chan</creator><creator>Park, Kwang-Hyun</creator><creator>Bak, In Seon</creator><creator>Song, Hyung-Nam</creator><creator>An, Yan</creator><creator>Lee, Su-Jin</creator><creator>Jung, Mira</creator><creator>Yoo, Kyeong-Won</creator><creator>Yu, Dae-Yeul</creator><creator>Kim, Yong-Sam</creator><creator>Oh, Byung-Ha</creator><creator>Woo, Eui-Jeon</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><scope>C6C</scope><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>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</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>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-6983-0270</orcidid><orcidid>https://orcid.org/0000-0002-6437-8470</orcidid></search><sort><creationdate>20190926</creationdate><title>In vivo genome editing using the Cpf1 ortholog derived from Eubacterium eligens</title><author>Ahn, Woo-Chan ; Park, Kwang-Hyun ; Bak, In Seon ; Song, Hyung-Nam ; An, Yan ; Lee, Su-Jin ; Jung, Mira ; Yoo, Kyeong-Won ; Yu, Dae-Yeul ; Kim, Yong-Sam ; Oh, Byung-Ha ; Woo, Eui-Jeon</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c474t-9914117503be4bbb3a8772757f8e74dd4be8433fb1e3596f904ec1b8ece31bba3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>38</topic><topic>38/70</topic><topic>42</topic><topic>42/41</topic><topic>631/1647/1511</topic><topic>631/61/17/1511</topic><topic>Animals</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>Blastocyst - metabolism</topic><topic>Blastocysts</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>Endonuclease</topic><topic>Endonucleases - genetics</topic><topic>Endonucleases - metabolism</topic><topic>Eubacterium - enzymology</topic><topic>Gene Editing - methods</topic><topic>Genome editing</topic><topic>Genomes</topic><topic>Humanities and Social Sciences</topic><topic>Immunodeficiency</topic><topic>Interleukin 2 receptors</topic><topic>Interleukin Receptor Common gamma Subunit - genetics</topic><topic>Interleukin Receptor Common gamma Subunit - metabolism</topic><topic>Magnesium - metabolism</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>multidisciplinary</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>RNA, Circular - genetics</topic><topic>Rodents</topic><topic>Science</topic><topic>Science (multidisciplinary)</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ahn, Woo-Chan</creatorcontrib><creatorcontrib>Park, Kwang-Hyun</creatorcontrib><creatorcontrib>Bak, In Seon</creatorcontrib><creatorcontrib>Song, Hyung-Nam</creatorcontrib><creatorcontrib>An, Yan</creatorcontrib><creatorcontrib>Lee, Su-Jin</creatorcontrib><creatorcontrib>Jung, Mira</creatorcontrib><creatorcontrib>Yoo, Kyeong-Won</creatorcontrib><creatorcontrib>Yu, Dae-Yeul</creatorcontrib><creatorcontrib>Kim, Yong-Sam</creatorcontrib><creatorcontrib>Oh, Byung-Ha</creatorcontrib><creatorcontrib>Woo, Eui-Jeon</creatorcontrib><collection>Springer Nature OA Free Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</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>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</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>Science Database</collection><collection>Biological Science Database</collection><collection>Publicly Available Content 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>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ahn, Woo-Chan</au><au>Park, Kwang-Hyun</au><au>Bak, In Seon</au><au>Song, Hyung-Nam</au><au>An, Yan</au><au>Lee, Su-Jin</au><au>Jung, Mira</au><au>Yoo, Kyeong-Won</au><au>Yu, Dae-Yeul</au><au>Kim, Yong-Sam</au><au>Oh, Byung-Ha</au><au>Woo, Eui-Jeon</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>In vivo genome editing using the Cpf1 ortholog derived from Eubacterium eligens</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2019-09-26</date><risdate>2019</risdate><volume>9</volume><issue>1</issue><spage>13911</spage><epage>7</epage><pages>13911-7</pages><artnum>13911</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>Cpf1 is an RNA-guided endonuclease that can be programmed to cleave DNA targets. Specific features, such as containing a short crRNA, creating a staggered cleavage pattern and having a low off-target rate, render Cpf1 a promising gene-editing tool. Here, we present a new Cpf1 ortholog, EeCpf1, as a genome-editing tool; this ortholog is derived from the gut bacterial species
Eubacterium eligens
. EeCpf1 exhibits a higher cleavage activity with the Mn
2+
metal cofactor and efficiently cuts the target DNA with an engineered, nucleotide extended crRNA at the 5′ target site. When mouse blastocysts were injected with multitargeting crRNAs against the IL2R-γ gene, an essential gene for immunodeficient mouse model production, EeCpf1 efficiently generated IL2R-γ knockout mice. For the first time, these results demonstrate that EeCpf1 can be used as an
in vivo
gene-editing tool for the production of knockout mice. The utilization of engineered crRNA with multiple target sites will help to explore the
in vivo
DNA cleavage activities of Cpf1 orthologs from other species that have not been demonstrated.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>31558757</pmid><doi>10.1038/s41598-019-50423-6</doi><tpages>7</tpages><orcidid>https://orcid.org/0000-0001-6983-0270</orcidid><orcidid>https://orcid.org/0000-0002-6437-8470</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | 38 38/70 42 42/41 631/1647/1511 631/61/17/1511 Animals Bacterial Proteins - genetics Bacterial Proteins - metabolism Blastocyst - metabolism Blastocysts Deoxyribonucleic acid DNA Endonuclease Endonucleases - genetics Endonucleases - metabolism Eubacterium - enzymology Gene Editing - methods Genome editing Genomes Humanities and Social Sciences Immunodeficiency Interleukin 2 receptors Interleukin Receptor Common gamma Subunit - genetics Interleukin Receptor Common gamma Subunit - metabolism Magnesium - metabolism Mice Mice, Inbred C57BL multidisciplinary Ribonucleic acid RNA RNA, Circular - genetics Rodents Science Science (multidisciplinary) |
| title | In vivo genome editing using the Cpf1 ortholog derived from Eubacterium eligens |
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