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Article ; Online: Active genetics comes alive: Exploring the broad applications of CRISPR-based selfish genetic elements (or gene-drives): Exploring the broad applications of CRISPR-based selfish genetic elements (or gene-drives).

Gantz, Valentino M / Bier, Ethan

BioEssays : news and reviews in molecular, cellular and developmental biology

2022 Volume 44, Issue 8, Page(s) e2100279

Abstract: Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based "active genetic" elements developed in 2015 bypassed the fundamental rules of traditional genetics. Inherited in a super-Mendelian fashion, such selfish genetic entities offered a ... ...

Abstract	Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based "active genetic" elements developed in 2015 bypassed the fundamental rules of traditional genetics. Inherited in a super-Mendelian fashion, such selfish genetic entities offered a variety of potential applications including: gene-drives to disseminate gene cassettes carrying desired traits throughout insect populations to control disease vectors or pest species, allelic drives biasing inheritance of preferred allelic variants, neutralizing genetic elements to delete and replace or to halt the spread of gene-drives, split-drives with the core constituent Cas9 endonuclease and guide RNA (gRNA) components inserted at separate genomic locations to accelerate assembly of complex arrays of genetic traits or to gain genetic entry into novel organisms (vertebrates, plants, bacteria), and interhomolog based copying systems in somatic cells to develop tools for treating inherited or infectious diseases. Here, we summarize the substantial advances that have been made on all of these fronts and look forward to the next phase of this rapidly expanding and impactful field.
MeSH term(s)	Animals ; CRISPR-Cas Systems/genetics ; Gene Drive Technology ; Gene Editing ; Inheritance Patterns ; RNA, Guide, CRISPR-Cas Systems/genetics
Chemical Substances	RNA, Guide, CRISPR-Cas Systems
Language	English
Publishing date	2022-06-09
Publishing country	United States
Document type	Journal Article ; Review ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	50140-2
ISSN	1521-1878 ; 0265-9247
ISSN (online)	1521-1878
ISSN	0265-9247
DOI	10.1002/bies.202100279
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Active genetics comes alive: Exploring the broad applications of CRISPR‐based selfish genetic elements (or gene‐drives): Exploring the broad applications of CRISPR‐based selfish genetic elements (or gene‐drives)

Gantz, Valentino M. / Bier, Ethan

BioEssays. 2022 Aug., v. 44, no. 8

2022

Abstract: Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)‐based “active genetic” elements developed in 2015 bypassed the fundamental rules of traditional genetics. Inherited in a super‐Mendelian fashion, such selfish genetic entities offered a ... ...

Abstract	Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)‐based “active genetic” elements developed in 2015 bypassed the fundamental rules of traditional genetics. Inherited in a super‐Mendelian fashion, such selfish genetic entities offered a variety of potential applications including: gene‐drives to disseminate gene cassettes carrying desired traits throughout insect populations to control disease vectors or pest species, allelic drives biasing inheritance of preferred allelic variants, neutralizing genetic elements to delete and replace or to halt the spread of gene‐drives, split‐drives with the core constituent Cas9 endonuclease and guide RNA (gRNA) components inserted at separate genomic locations to accelerate assembly of complex arrays of genetic traits or to gain genetic entry into novel organisms (vertebrates, plants, bacteria), and interhomolog based copying systems in somatic cells to develop tools for treating inherited or infectious diseases. Here, we summarize the substantial advances that have been made on all of these fronts and look forward to the next phase of this rapidly expanding and impactful field.
Keywords	CRISPR-associated endonuclease 9 ; RNA ; genes ; genomics ; insects ; pests
Language	English
Dates of publication	2022-08
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	REVIEW
ZDB-ID	50140-2
ISSN	1521-1878 ; 0265-9247
ISSN (online)	1521-1878
ISSN	0265-9247
DOI	10.1002/bies.202100279
Database	NAL-Catalogue (AGRICOLA)

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Article: CRISPR-based gene drives generate super-Mendelian inheritance in the disease vector

Harvey-Samuel, Tim / Feng, Xuechun / Okamoto, Emily M / Purusothaman, Deepak-Kumar / Leftwich, Philip T / Alphey, Luke / Gantz, Valentino M

bioRxiv : the preprint server for biology

2023

Abstract: ... ...

Abstract	Culex
Language	English
Publishing date	2023-06-12
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.06.12.544656
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: A nickase Cas9 gene-drive system promotes super-Mendelian inheritance in Drosophila.

López Del Amo, Víctor / Juste, Sara Sanz / Gantz, Valentino M

Cell reports

2022 Volume 39, Issue 8, Page(s) 110843

Abstract: CRISPR-based gene-drives have been proposed for managing insect populations, including disease-transmitting mosquitoes, due to their ability to bias their inheritance toward super-Mendelian rates (>50%). Current technologies use a Cas9 that introduces ... ...

Abstract	CRISPR-based gene-drives have been proposed for managing insect populations, including disease-transmitting mosquitoes, due to their ability to bias their inheritance toward super-Mendelian rates (>50%). Current technologies use a Cas9 that introduces DNA double-strand breaks into the opposing wild-type allele to replace it with a copy of the gene-drive allele via DNA homology-directed repair. However, the use of different Cas9 versions is unexplored, and alternative approaches could increase the available toolkit for gene-drive designs. Here, we report a gene-drive that relies on Cas9 nickases that generate staggered paired nicks in DNA to propagate the engineered gene-drive cassette. We show that generating 5' overhangs in the system yields efficient allelic conversion. The nickase gene-drive arrangement produces large, stereotyped deletions that are advantageous to eliminate viable animals carrying small mutations when targeting essential genes. Our nickase approach should expand the repertoire for gene-drive arrangements aimed at applications in mosquitoes and beyond.
MeSH term(s)	Animals ; CRISPR-Cas Systems/genetics ; DNA ; Deoxyribonuclease I/metabolism ; Drosophila/metabolism ; Gene Drive Technology ; Gene Editing
Chemical Substances	DNA (9007-49-2) ; Deoxyribonuclease I (EC 3.1.21.1)
Language	English
Publishing date	2022-05-22
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2649101-1
ISSN	2211-1247 ; 2211-1247
ISSN (online)	2211-1247
ISSN	2211-1247
DOI	10.1016/j.celrep.2022.110843
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Targeting double-strand break indel byproducts with secondary guide RNAs improves Cas9 HDR-mediated genome editing efficiencies.

Bodai, Zsolt / Bishop, Alena L / Gantz, Valentino M / Komor, Alexis C

Nature communications

2022 Volume 13, Issue 1, Page(s) 2351

Abstract: Programmable double-strand DNA breaks (DSBs) can be harnessed for precision genome editing through manipulation of the homology-directed repair (HDR) pathway. However, end-joining repair pathways often outcompete HDR and introduce insertions and ... ...

Abstract	Programmable double-strand DNA breaks (DSBs) can be harnessed for precision genome editing through manipulation of the homology-directed repair (HDR) pathway. However, end-joining repair pathways often outcompete HDR and introduce insertions and deletions of bases (indels) at the DSB site, decreasing precision outcomes. It has been shown that indel sequences for a given DSB site are reproducible and can even be predicted. Here, we report a general strategy (the "double tap" method) to improve HDR-mediated precision genome editing efficiencies that takes advantage of the reproducible nature of indel sequences. The method simply involves the use of multiple gRNAs: a primary gRNA that targets the wild-type genomic sequence, and one or more secondary gRNAs that target the most common indel sequence(s), which in effect provides a "second chance" at HDR-mediated editing. This proof-of-principle study presents the double tap method as a simple yet effective option for enhancing precision editing in mammalian cells.
MeSH term(s)	Animals ; CRISPR-Cas Systems/genetics ; DNA End-Joining Repair ; Gene Editing/methods ; Mammals/genetics ; Recombinational DNA Repair ; RNA, Guide, CRISPR-Cas Systems
Language	English
Publishing date	2022-05-09
Publishing country	England
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-022-29989-9
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: CRISPR-based gene drives generate super-Mendelian inheritance in the disease vector Culex quinquefasciatus.

Harvey-Samuel, Tim / Feng, Xuechun / Okamoto, Emily M / Purusothaman, Deepak-Kumar / Leftwich, Philip T / Alphey, Luke / Gantz, Valentino M

Nature communications

2023 Volume 14, Issue 1, Page(s) 7561

Abstract: Culex mosquitoes pose a significant public health threat as vectors for a variety of diseases including West Nile virus and lymphatic filariasis, and transmit pathogens threatening livestock, companion animals, and endangered birds. Rampant insecticide ... ...

Abstract	Culex mosquitoes pose a significant public health threat as vectors for a variety of diseases including West Nile virus and lymphatic filariasis, and transmit pathogens threatening livestock, companion animals, and endangered birds. Rampant insecticide resistance makes controlling these mosquitoes challenging and necessitates the development of new control strategies. Gene drive technologies have made significant progress in other mosquito species, although similar advances have been lagging in Culex. Here we test a CRISPR-based homing gene drive for Culex quinquefasciatus, and show that the inheritance of two split-gene-drive transgenes, targeting different loci, are biased in the presence of a Cas9-expressing transgene although with modest efficiencies. Our findings extend the list of disease vectors where engineered homing gene drives have been demonstrated to include Culex alongside Anopheles and Aedes, and pave the way for future development of these technologies to control Culex mosquitoes.
MeSH term(s)	Animals ; Culex/genetics ; Gene Drive Technology ; Mosquito Vectors/genetics ; Aedes/genetics ; Disease Vectors
Language	English
Publishing date	2023-11-20
Publishing country	England
Document type	Journal Article
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-023-41834-1
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Gene editing technologies and applications for insects.

Gantz, Valentino M / Akbari, Omar S

Current opinion in insect science

2018 Volume 28, Page(s) 66–72

Abstract: Initially discovered in bacteria, CRISPR-based genome editing endonucleases have proven remarkably amenable for adaptation to insects. To date, these endonucleases have been utilized in a plethora of both model and non-model insects including diverse ... ...

Abstract	Initially discovered in bacteria, CRISPR-based genome editing endonucleases have proven remarkably amenable for adaptation to insects. To date, these endonucleases have been utilized in a plethora of both model and non-model insects including diverse flies, bees, beetles, butterflies, moths, and grasshoppers, to name a few, thereby revolutionizing functional genomics of insects. In addition to basic genome editing, they have also been invaluable for advanced genome engineering and synthetic biology applications. Here we explore the recent genome editing advancements in insects for generating site-specific genomic mutations, insertions, deletions, as well as more advanced applications such as Homology Assisted Genome Knock-in (HACK), potential to utilize DNA base editing, generating predictable reciprocal chromosomal translocations, and development gene drives to control the fate of wild populations.
MeSH term(s)	Animals ; Gene Deletion ; Gene Drive Technology/methods ; Gene Editing/instrumentation ; Gene Editing/methods ; Gene Knock-In Techniques/methods ; Genome ; Insecta/genetics ; Mutagenesis, Insertional ; Mutation ; Translocation, Genetic
Language	English
Publishing date	2018-05-22
Publishing country	Netherlands
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Review
ZDB-ID	2772833-X
ISSN	2214-5753 ; 2214-5745
ISSN (online)	2214-5753
ISSN	2214-5745
DOI	10.1016/j.cois.2018.05.006
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Targeting double-strand break indel byproducts with secondary guide RNAs improves Cas9 HDR-mediated genome editing efficiencies

Zsolt Bodai / Alena L. Bishop / Valentino M. Gantz / Alexis C. Komor

Nature Communications, Vol 13, Iss 1, Pp 1-

2022 Volume 15

Abstract: Programmable double-strand DNA breaks (DSBs) can be harnessed for precision genome editing through manipulation of the homology-directed repair (HDR) pathway. Here the authors report the development of the double tap - double tap implements secondary ... ...

Abstract	Programmable double-strand DNA breaks (DSBs) can be harnessed for precision genome editing through manipulation of the homology-directed repair (HDR) pathway. Here the authors report the development of the double tap - double tap implements secondary gRNAs which target Cas9 to common indel sequences and provides a second chance at HDR.
Keywords	Science ; Q
Language	English
Publishing date	2022-05-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: The dawn of active genetics.

Gantz, Valentino M / Bier, Ethan

BioEssays : news and reviews in molecular, cellular and developmental biology

2016 Volume 38, Issue 1, Page(s) 50–63

Abstract: On December 18, 2014, a yellow female fly quietly emerged from her pupal case. What made her unique was that she had only one parent carrying a mutant allele of this classic recessive locus. Then, one generation later, after mating with a wild-type male, ...

Abstract	On December 18, 2014, a yellow female fly quietly emerged from her pupal case. What made her unique was that she had only one parent carrying a mutant allele of this classic recessive locus. Then, one generation later, after mating with a wild-type male, all her offspring displayed the same recessive yellow phenotype. Further analysis of other such yellow females revealed that the construct causing the mutation was converting the opposing chromosome with 95% efficiency. These simple results, seen also in mosquitoes and yeast, open the door to a new era of genetics wherein the laws of traditional Mendelian inheritance can be bypassed for a broad variety of purposes. Here, we consider the implications of this fundamentally new form of "active genetics," its applications for gene drives, reversal and amplification strategies, its potential for contributing to cell and gene therapy strategies, and ethical/biosafety considerations associated with such active genetic elements. Also watch the Video Abstract.
MeSH term(s)	Alleles ; Animals ; Animals, Genetically Modified/genetics ; CRISPR-Cas Systems/genetics ; Chromosomes/genetics ; Drosophila melanogaster/genetics ; Female ; Gene Editing ; Genes, Recessive ; Male ; Mutation/genetics ; Phenotype
Language	English
Publishing date	2016-01
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	50140-2
ISSN	1521-1878 ; 0265-9247
ISSN (online)	1521-1878
ISSN	0265-9247
DOI	10.1002/bies.201500102
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Double-tap gene drive uses iterative genome targeting to help overcome resistance alleles.

Bishop, Alena L / López Del Amo, Víctor / Okamoto, Emily M / Bodai, Zsolt / Komor, Alexis C / Gantz, Valentino M

Nature communications

2022 Volume 13, Issue 1, Page(s) 2595

Abstract: Homing CRISPR gene drives could aid in curbing the spread of vector-borne diseases and controlling crop pest and invasive species populations due to an inheritance rate that surpasses Mendelian laws. However, this technology suffers from resistance ... ...

Abstract	Homing CRISPR gene drives could aid in curbing the spread of vector-borne diseases and controlling crop pest and invasive species populations due to an inheritance rate that surpasses Mendelian laws. However, this technology suffers from resistance alleles formed when the drive-induced DNA break is repaired by error-prone pathways, which creates mutations that disrupt the gRNA recognition sequence and prevent further gene-drive propagation. Here, we attempt to counteract this by encoding additional gRNAs that target the most commonly generated resistance alleles into the gene drive, allowing a second opportunity at gene-drive conversion. Our presented "double-tap" strategy improved drive efficiency by recycling resistance alleles. The double-tap drive also efficiently spreads in caged populations, outperforming the control drive. Overall, this double-tap strategy can be readily implemented in any CRISPR-based gene drive to improve performance, and similar approaches could benefit other systems suffering from low HDR frequencies, such as mammalian cells or mouse germline transformations.
MeSH term(s)	Alleles ; Animals ; CRISPR-Cas Systems/genetics ; Gene Drive Technology ; Germ Cells ; Mammals/genetics ; Mice ; RNA, Guide, CRISPR-Cas Systems/genetics
Chemical Substances	RNA, Guide, CRISPR-Cas Systems
Language	English
Publishing date	2022-05-09
Publishing country	England
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-022-29868-3
Database	MEDical Literature Analysis and Retrieval System OnLINE

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