Repeat mediated excision of gene drive elements for restoring wild-type populations.
Here, we demonstrate that single strand annealing (SSA) can be co-opted for the precise autocatalytic excision of a drive element. We have termed this technology Repeat Mediated Excision of a Drive Element (ReMEDE). By engineering direct repeats flanking the drive allele and inducing a double-strand...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2024-11-01
|
| Series: | PLoS Genetics |
| Online Access: | https://doi.org/10.1371/journal.pgen.1011450 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850075323941519360 |
|---|---|
| author | Pratima R Chennuri Josef Zapletal Raquel D Monfardini Martial Loth Ndeffo-Mbah Zach N Adelman Kevin M Myles |
| author_facet | Pratima R Chennuri Josef Zapletal Raquel D Monfardini Martial Loth Ndeffo-Mbah Zach N Adelman Kevin M Myles |
| author_sort | Pratima R Chennuri |
| collection | DOAJ |
| description | Here, we demonstrate that single strand annealing (SSA) can be co-opted for the precise autocatalytic excision of a drive element. We have termed this technology Repeat Mediated Excision of a Drive Element (ReMEDE). By engineering direct repeats flanking the drive allele and inducing a double-strand DNA break (DSB) at a second endonuclease target site within the allele, we increased the utilization of SSA repair. ReMEDE was incorporated into the mutagenic chain reaction (MCR) gene drive targeting the yellow gene of Drosophila melanogaster, successfully replacing drive alleles with wild-type alleles. Sequencing across the Cas9 target site confirmed transgene excision by SSA after pair-mated outcrosses with yReMEDE females, revealing ~4% inheritance of an engineered silent TcG marker sequence. However, phenotypically wild-type flies with alleles of indeterminate biogenesis also were observed, retaining the TGG sequence (~16%) or harboring a silent gGG mutation (~0.5%) at the PAM site. Additionally, ~14% of alleles in the F2 flies were intact or uncut paternally inherited alleles, indicating limited maternal deposition of Cas9 RNP. Although ReMEDE requires further research and development, the technology has some promising features as a gene drive mitigation strategy, notably its potential to restore wild-type populations without additional transgenic releases or large-scale environmental modifications. |
| format | Article |
| id | doaj-art-cb7a7f2ee10b482ba11c9784c5cda4b3 |
| institution | DOAJ |
| issn | 1553-7390 1553-7404 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Genetics |
| spelling | doaj-art-cb7a7f2ee10b482ba11c9784c5cda4b32025-08-20T02:46:20ZengPublic Library of Science (PLoS)PLoS Genetics1553-73901553-74042024-11-012011e101145010.1371/journal.pgen.1011450Repeat mediated excision of gene drive elements for restoring wild-type populations.Pratima R ChennuriJosef ZapletalRaquel D MonfardiniMartial Loth Ndeffo-MbahZach N AdelmanKevin M MylesHere, we demonstrate that single strand annealing (SSA) can be co-opted for the precise autocatalytic excision of a drive element. We have termed this technology Repeat Mediated Excision of a Drive Element (ReMEDE). By engineering direct repeats flanking the drive allele and inducing a double-strand DNA break (DSB) at a second endonuclease target site within the allele, we increased the utilization of SSA repair. ReMEDE was incorporated into the mutagenic chain reaction (MCR) gene drive targeting the yellow gene of Drosophila melanogaster, successfully replacing drive alleles with wild-type alleles. Sequencing across the Cas9 target site confirmed transgene excision by SSA after pair-mated outcrosses with yReMEDE females, revealing ~4% inheritance of an engineered silent TcG marker sequence. However, phenotypically wild-type flies with alleles of indeterminate biogenesis also were observed, retaining the TGG sequence (~16%) or harboring a silent gGG mutation (~0.5%) at the PAM site. Additionally, ~14% of alleles in the F2 flies were intact or uncut paternally inherited alleles, indicating limited maternal deposition of Cas9 RNP. Although ReMEDE requires further research and development, the technology has some promising features as a gene drive mitigation strategy, notably its potential to restore wild-type populations without additional transgenic releases or large-scale environmental modifications.https://doi.org/10.1371/journal.pgen.1011450 |
| spellingShingle | Pratima R Chennuri Josef Zapletal Raquel D Monfardini Martial Loth Ndeffo-Mbah Zach N Adelman Kevin M Myles Repeat mediated excision of gene drive elements for restoring wild-type populations. PLoS Genetics |
| title | Repeat mediated excision of gene drive elements for restoring wild-type populations. |
| title_full | Repeat mediated excision of gene drive elements for restoring wild-type populations. |
| title_fullStr | Repeat mediated excision of gene drive elements for restoring wild-type populations. |
| title_full_unstemmed | Repeat mediated excision of gene drive elements for restoring wild-type populations. |
| title_short | Repeat mediated excision of gene drive elements for restoring wild-type populations. |
| title_sort | repeat mediated excision of gene drive elements for restoring wild type populations |
| url | https://doi.org/10.1371/journal.pgen.1011450 |
| work_keys_str_mv | AT pratimarchennuri repeatmediatedexcisionofgenedriveelementsforrestoringwildtypepopulations AT josefzapletal repeatmediatedexcisionofgenedriveelementsforrestoringwildtypepopulations AT raqueldmonfardini repeatmediatedexcisionofgenedriveelementsforrestoringwildtypepopulations AT martiallothndeffombah repeatmediatedexcisionofgenedriveelementsforrestoringwildtypepopulations AT zachnadelman repeatmediatedexcisionofgenedriveelementsforrestoringwildtypepopulations AT kevinmmyles repeatmediatedexcisionofgenedriveelementsforrestoringwildtypepopulations |