Creation of targeted genomic deletions using TALEN or CRISPR/Cas nuclease pairs in one‐cell mouse embryos
The use of TALEN and CRISPR/CAS nucleases is becoming increasingly popular as a means to edit single target sites in one‐cell mouse embryos. Nevertheless, an area that has received less attention concerns the engineering of structural genome variants and the necessary religation of two distant doubl...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2015-01-01
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| Series: | FEBS Open Bio |
| Subjects: | |
| Online Access: | https://doi.org/10.1016/j.fob.2014.11.009 |
| _version_ | 1811178370618097664 |
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| author | Christina Brandl Oskar Ortiz Bernhard Röttig Benedikt Wefers Wolfgang Wurst Ralf Kühn |
| author_facet | Christina Brandl Oskar Ortiz Bernhard Röttig Benedikt Wefers Wolfgang Wurst Ralf Kühn |
| author_sort | Christina Brandl |
| collection | DOAJ |
| description | The use of TALEN and CRISPR/CAS nucleases is becoming increasingly popular as a means to edit single target sites in one‐cell mouse embryos. Nevertheless, an area that has received less attention concerns the engineering of structural genome variants and the necessary religation of two distant double‐strand breaks. Herein, we applied pairs of TALEN or sgRNAs and Cas9 to create deletions in theRab38 gene. We found that the deletion of 3.2 or 9.3 kb, but not of 30 kb, occurs at a frequency of 6–37%. This is sufficient for the direct production of mutants by embryo microinjection. Therefore, deletions up to ∼10 kb can be readily achieved for modeling human disease alleles. This work represents an important step towards the establishment of new protocols that support the ligation of remote DSB ends to achieve even larger rearrangements. |
| first_indexed | 2024-04-11T06:18:11Z |
| format | Article |
| id | doaj.art-1b42b524856e4b2697264d7f9efd8c2e |
| institution | Directory Open Access Journal |
| issn | 2211-5463 |
| language | English |
| last_indexed | 2024-04-11T06:18:11Z |
| publishDate | 2015-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj.art-1b42b524856e4b2697264d7f9efd8c2e2022-12-22T04:40:59ZengWileyFEBS Open Bio2211-54632015-01-0151263510.1016/j.fob.2014.11.009Creation of targeted genomic deletions using TALEN or CRISPR/Cas nuclease pairs in one‐cell mouse embryosChristina Brandl0Oskar Ortiz1Bernhard Röttig2Benedikt Wefers3Wolfgang Wurst4Ralf Kühn5Institute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Neuherberg, GermanyInstitute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Neuherberg, GermanyInstitute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Neuherberg, GermanyInstitute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Neuherberg, GermanyInstitute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Neuherberg, GermanyInstitute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Neuherberg, GermanyThe use of TALEN and CRISPR/CAS nucleases is becoming increasingly popular as a means to edit single target sites in one‐cell mouse embryos. Nevertheless, an area that has received less attention concerns the engineering of structural genome variants and the necessary religation of two distant double‐strand breaks. Herein, we applied pairs of TALEN or sgRNAs and Cas9 to create deletions in theRab38 gene. We found that the deletion of 3.2 or 9.3 kb, but not of 30 kb, occurs at a frequency of 6–37%. This is sufficient for the direct production of mutants by embryo microinjection. Therefore, deletions up to ∼10 kb can be readily achieved for modeling human disease alleles. This work represents an important step towards the establishment of new protocols that support the ligation of remote DSB ends to achieve even larger rearrangements.https://doi.org/10.1016/j.fob.2014.11.009TALENsDisease modelOne-cell embryoMouse mutantCRISPRCas9 |
| spellingShingle | Christina Brandl Oskar Ortiz Bernhard Röttig Benedikt Wefers Wolfgang Wurst Ralf Kühn Creation of targeted genomic deletions using TALEN or CRISPR/Cas nuclease pairs in one‐cell mouse embryos FEBS Open Bio TALENs Disease model One-cell embryo Mouse mutant CRISPR Cas9 |
| title | Creation of targeted genomic deletions using TALEN or CRISPR/Cas nuclease pairs in one‐cell mouse embryos |
| title_full | Creation of targeted genomic deletions using TALEN or CRISPR/Cas nuclease pairs in one‐cell mouse embryos |
| title_fullStr | Creation of targeted genomic deletions using TALEN or CRISPR/Cas nuclease pairs in one‐cell mouse embryos |
| title_full_unstemmed | Creation of targeted genomic deletions using TALEN or CRISPR/Cas nuclease pairs in one‐cell mouse embryos |
| title_short | Creation of targeted genomic deletions using TALEN or CRISPR/Cas nuclease pairs in one‐cell mouse embryos |
| title_sort | creation of targeted genomic deletions using talen or crispr cas nuclease pairs in one cell mouse embryos |
| topic | TALENs Disease model One-cell embryo Mouse mutant CRISPR Cas9 |
| url | https://doi.org/10.1016/j.fob.2014.11.009 |
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