Efficient gene targeting in Drosophila by direct embryo injection with zinc-finger nucleases.
We report very high gene targeting frequencies in Drosophila by direct embryo injection of mRNAs encoding specific zinc-finger nucleases (ZFNs). Both local mutagenesis via nonhomologous end joining (NHEJ) and targeted gene replacement via homologous recombination (HR) have been achieved in up to 10%...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Journal article |
| Language: | English |
| Published: |
2008
|
| _version_ | 1826291727740174336 |
|---|---|
| author | Beumer, K Trautman, J Bozas, A Liu, J Rutter, J Gall, J Carroll, D |
| author_facet | Beumer, K Trautman, J Bozas, A Liu, J Rutter, J Gall, J Carroll, D |
| author_sort | Beumer, K |
| collection | OXFORD |
| description | We report very high gene targeting frequencies in Drosophila by direct embryo injection of mRNAs encoding specific zinc-finger nucleases (ZFNs). Both local mutagenesis via nonhomologous end joining (NHEJ) and targeted gene replacement via homologous recombination (HR) have been achieved in up to 10% of all targets at a given locus. In embryos that are wild type for DNA repair, the products are dominated by NHEJ mutations. In recipients deficient in the NHEJ component, DNA ligase IV, the majority of products arise by HR with a coinjected donor DNA, with no loss of overall efficiency in target modification. We describe the application of the ZFN injection procedure to mutagenesis by NHEJ of 2 new genes in Drosophila melanogaster: coil and pask. Pairs of novel ZFNs designed for targets within those genes led to the production of null mutations at each locus. The injection procedure is much more rapid than earlier approaches and makes possible the generation and recovery of targeted gene alterations at essentially any locus within 2 fly generations. |
| first_indexed | 2024-03-07T03:03:46Z |
| format | Journal article |
| id | oxford-uuid:b1d4395c-415a-4a88-8840-5d08a8183537 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T03:03:46Z |
| publishDate | 2008 |
| record_format | dspace |
| spelling | oxford-uuid:b1d4395c-415a-4a88-8840-5d08a81835372022-03-27T04:07:02ZEfficient gene targeting in Drosophila by direct embryo injection with zinc-finger nucleases.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:b1d4395c-415a-4a88-8840-5d08a8183537EnglishSymplectic Elements at Oxford2008Beumer, KTrautman, JBozas, ALiu, JRutter, JGall, JCarroll, DWe report very high gene targeting frequencies in Drosophila by direct embryo injection of mRNAs encoding specific zinc-finger nucleases (ZFNs). Both local mutagenesis via nonhomologous end joining (NHEJ) and targeted gene replacement via homologous recombination (HR) have been achieved in up to 10% of all targets at a given locus. In embryos that are wild type for DNA repair, the products are dominated by NHEJ mutations. In recipients deficient in the NHEJ component, DNA ligase IV, the majority of products arise by HR with a coinjected donor DNA, with no loss of overall efficiency in target modification. We describe the application of the ZFN injection procedure to mutagenesis by NHEJ of 2 new genes in Drosophila melanogaster: coil and pask. Pairs of novel ZFNs designed for targets within those genes led to the production of null mutations at each locus. The injection procedure is much more rapid than earlier approaches and makes possible the generation and recovery of targeted gene alterations at essentially any locus within 2 fly generations. |
| spellingShingle | Beumer, K Trautman, J Bozas, A Liu, J Rutter, J Gall, J Carroll, D Efficient gene targeting in Drosophila by direct embryo injection with zinc-finger nucleases. |
| title | Efficient gene targeting in Drosophila by direct embryo injection with zinc-finger nucleases. |
| title_full | Efficient gene targeting in Drosophila by direct embryo injection with zinc-finger nucleases. |
| title_fullStr | Efficient gene targeting in Drosophila by direct embryo injection with zinc-finger nucleases. |
| title_full_unstemmed | Efficient gene targeting in Drosophila by direct embryo injection with zinc-finger nucleases. |
| title_short | Efficient gene targeting in Drosophila by direct embryo injection with zinc-finger nucleases. |
| title_sort | efficient gene targeting in drosophila by direct embryo injection with zinc finger nucleases |
| work_keys_str_mv | AT beumerk efficientgenetargetingindrosophilabydirectembryoinjectionwithzincfingernucleases AT trautmanj efficientgenetargetingindrosophilabydirectembryoinjectionwithzincfingernucleases AT bozasa efficientgenetargetingindrosophilabydirectembryoinjectionwithzincfingernucleases AT liuj efficientgenetargetingindrosophilabydirectembryoinjectionwithzincfingernucleases AT rutterj efficientgenetargetingindrosophilabydirectembryoinjectionwithzincfingernucleases AT gallj efficientgenetargetingindrosophilabydirectembryoinjectionwithzincfingernucleases AT carrolld efficientgenetargetingindrosophilabydirectembryoinjectionwithzincfingernucleases |