(14) Methods for sex-sorting insects - W)2024091883A2
Provided herein are methods of sex-sorting a plurality of insects based on sex-specific gene expression, the method comprising (a) generating an exogenous nucleic acid molecule; (b) delivering the exogenous nucleic acid molecule into an insect from the plurality of insects, wherein the exogenous nucleic acid molecule comprises a promoter region, a sex-specific splicing module, a reporter gene, and a transcription terminator; (c) detecting sex-specific gene expression of the reporter gene; and (d) sorting the insect from the plurality of insects based on the detecting of the sex-specific gene expression in step (c), thereby sex-sorting the insect based on the sex-specific gene expression.
Provided herein are methods of sex-sorting a plurality of insects based on sex-specific gene expression, the method comprising (a) generating an exogenous nucleic acid molecule; (b) delivering the exogenous nucleic acid molecule into an insect from the plurality of insects, wherein the exogenous nucleic acid molecule comprises a promoter region, a sex-specific splicing module, a reporter gene, and a transcription terminator; (c) detecting sex-specific gene expression of the reporter gene; and (d) sorting the insect from the plurality of insects based on the detecting of the sex-specific gene expression in step (c), thereby sex-sorting the insect based on the sex-specific gene expression.
(13) Confinable population suppression system - WO2023230572A1
Provided herein are methods and transgenic systems termed Ifegenia (Inherited Female Elimination by Genetically Encoded Nucleases to Interrupt Alleles) comprising transgenic animal strains encoding Cas9 and/or gRNA that targets a female essential gene which are capable of passing down these genes as well as mutant female essential genes in wild populations in order to suppress the population of the animals, as well as methods and systems for making such animals. In some instances, the methods and systems provided herein result in both somatic and heritable germline mutations of the female essential gene resulting in daughter killing, and female essential gene mutant males reproductively viable to pass along the female essential gene mutation and related transgenes into subsequent generations. The methods and systems are adaptable to population control of insects, in particular mosquitoes such as Anopheles gambiae.
Provided herein are methods and transgenic systems termed Ifegenia (Inherited Female Elimination by Genetically Encoded Nucleases to Interrupt Alleles) comprising transgenic animal strains encoding Cas9 and/or gRNA that targets a female essential gene which are capable of passing down these genes as well as mutant female essential genes in wild populations in order to suppress the population of the animals, as well as methods and systems for making such animals. In some instances, the methods and systems provided herein result in both somatic and heritable germline mutations of the female essential gene resulting in daughter killing, and female essential gene mutant males reproductively viable to pass along the female essential gene mutation and related transgenes into subsequent generations. The methods and systems are adaptable to population control of insects, in particular mosquitoes such as Anopheles gambiae.
(12) Systems and methods for identifying novel crispr associated proteins
WO WO2022109466A1 Omar AKBARI The Regents Of The University Of California
Priority 2020-11-23 • Filed 2021-11-23 • Published 2022-05-27
Provided herein are systems and methods for identifying Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated proteins. For example, a method of identifying Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated proteins can include: (a) obtaining a …
WO WO2022109466A1 Omar AKBARI The Regents Of The University Of California
Priority 2020-11-23 • Filed 2021-11-23 • Published 2022-05-27
Provided herein are systems and methods for identifying Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated proteins. For example, a method of identifying Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated proteins can include: (a) obtaining a …
(11) Engineered reproductive isolation in animals
WO WO2021262794A1 Omar AKBARI The Regents Of The University Of California
Priority 2020-06-23 • Filed 2021-06-23 • Published 2021-12-30
Provided herein are methods of engineering a reproductive barrier in an insect including (a) introducing into a first insect a first nucleic acid sequence, where the first nucleic acid sequence targets a genomic sequence, where the genomic sequence is proximal to a transcription start site of a …
WO WO2021262794A1 Omar AKBARI The Regents Of The University Of California
Priority 2020-06-23 • Filed 2021-06-23 • Published 2021-12-30
Provided herein are methods of engineering a reproductive barrier in an insect including (a) introducing into a first insect a first nucleic acid sequence, where the first nucleic acid sequence targets a genomic sequence, where the genomic sequence is proximal to a transcription start site of a …
(10) One-locus inducible precision guided sterile insect technique or temperature- …
WO WO2021242782A1 Omar S. AKBARI The Regents Of The University Of California
Priority 2020-05-26 • Filed 2021-05-25 • Published 2021-12-02
Provided is a gene editing system comprising (i) a polynucleotide encoding an endonuclease under the control of an inducible expressing regulatory sequence; (ii) a guide polynucleotide targeting a female-essential genomic sequence that is required for female-specific viability, and (iii) a guide …
WO WO2021242782A1 Omar S. AKBARI The Regents Of The University Of California
Priority 2020-05-26 • Filed 2021-05-25 • Published 2021-12-02
Provided is a gene editing system comprising (i) a polynucleotide encoding an endonuclease under the control of an inducible expressing regulatory sequence; (ii) a guide polynucleotide targeting a female-essential genomic sequence that is required for female-specific viability, and (iii) a guide …
(9) An ultrasensitive rapid and portable case13d-based diagnostic assay
WO WO2021226278A1 Omar AKBARI The Regents Of The University Of California
Priority 2020-05-06 • Filed 2021-05-05 • Published 2021-11-11
Provided herein is a viral RNA detection system, utilizing the RNA-targeting properties of the optimized Casl3d enzyme, CasRx, to detect SARS-CoV-2 RNA, e.g., synthetic SARS- CoV-2 RNA. The system detects novel target sequences conserved within the actively evolving genome, to provide a panel of …
WO WO2021226278A1 Omar AKBARI The Regents Of The University Of California
Priority 2020-05-06 • Filed 2021-05-05 • Published 2021-11-11
Provided herein is a viral RNA detection system, utilizing the RNA-targeting properties of the optimized Casl3d enzyme, CasRx, to detect SARS-CoV-2 RNA, e.g., synthetic SARS- CoV-2 RNA. The system detects novel target sequences conserved within the actively evolving genome, to provide a panel of …
(8) Methods and compositions for sexing and sterilization in drosophila suzukii and Aedes aegypti Provided herein is a next-generation highly-efficient technology that can be used for biocontrol of D . suzukii and/or Aedes aegypti. The composition and technique termed precision guided SIT (pgSIT) functions by exploiting the precision and accuracy of CRISPR to simultaneously disrupt genes essential for either female viability or male fertility. It utilizes a simple breeding scheme requiring two homozygous strains - one expressing Cas9 and the other expressing double guide RNAs (dgRNAs). A single mating between these strains mechanistically results in synchro¬ nous RNA-guided dominant biallelic knockouts of both target genes throughout development, resulting in the complete penetrance of desired phenotypes in all progeny. This document provides methods and compositions relating to producing such insect eggs, insects, insect populations and uses thereof in reducing a wild-type in¬ sect population, along with methods and materials for producing genetically modified progeny of D . suzukii and/or Aedes aegypti.
(7) RNA Targeting Cas Enzymes
Provided herein are compositions and methods for CRISPR based RNA-targeting. The compositions include nucleic acid molecules comprising a sequence encoding a Cas13 polypeptide and a sequence encoding a guide RNA comprising one or more spacers and one or more Cas13-specific direct repeats, where the spacers are capable of specifically hybridizing with one or more target RNAs. The disclosure further provides methods of modifying a target RNA in a cell and transgenic organisms.
Provided herein are compositions and methods for CRISPR based RNA-targeting. The compositions include nucleic acid molecules comprising a sequence encoding a Cas13 polypeptide and a sequence encoding a guide RNA comprising one or more spacers and one or more Cas13-specific direct repeats, where the spacers are capable of specifically hybridizing with one or more target RNAs. The disclosure further provides methods of modifying a target RNA in a cell and transgenic organisms.
(6) ENDONUCLEASE SEXING AND STERILIZING IN INSECTS
- University of California, Riverside
- Filed: 11/2017
- Patent Application No. PCT/US2018/061886
- Abstract: Methods of the disclosed precision guided sterile insect technique (pgSIT) include methods for directing male sexing in a genetically modified insect and methods of producing a progeny of genetically modified sterile male insect egg. These methods include integrating at least one nucleic acid sequence into a genome of a first insect, the at least one nucleic acid sequence having at least one first guide polynucleotide targeting a female-essential genomic sequence that is required for female-specific viability, introducing an endonuclease into a second insect, and genetically crossing the first insect and the second insect thereby producing progeny expressing the endonuclease and the at least one nucleic acid sequence. For male sterility a second guide polynucleotide targets a male sterility genomic sequence that is required for male-specific sterility.
(5) ENGINEERED RESISTANCE TO VIRUSES IN TRANSGENIC MOSQUITOES
- Abstract: Recent Zika virus (ZIKV) outbreaks have highlighted the need for novel strategies to control arboviral transmission. Here, we describe the development of the first mosquitoes synthetically engineered to be 100% resistant to ZIKV. We show that a synthetic polycistronic cluster of engineered microRNAs (miRNAs) targeting ZIKV (termed the anti-ZIKV transgene) is expressed and fully processed following a blood meal in Aedes aegypti (Ae. aegypti), ensuring the formation of mature synthetic miRNAs in the midgut lumen where ZIKV resides in the early stages of infection. Critically, we demonstrate that homozygous Ae. aegypti mosquitoes expressing the anti-ZIKV transgene are unable to be infected with, and cannot transmit, ZIKV. Together, these compelling results provide a promising path forward for development of effective genetic-based ZIKV control strategies, which could potentially be extended to control other arboviruses.
(3) POPULATION CONTROL USING ENGINEERED TRANSLOCATIONS
- Publication number: 20160345556
- Abstract: Embodiments provided herein relate to systems for synthetically-engineered reciprocal chromosomal translocation for gene insertion into a population of organisms such as insects. pdf
- Type: Application
- Filed: May 25, 2016
- Publication date: December 1, 2016
- Inventors: Bruce A. Hay, Omar S. Akbari, Anna B. Buchman
(2) REGULATION OF INSECT POPULATIONS
- Publication number: 20150237838
- Abstract: The present technology generally relates to compositions, methods, and populations that allow for the regulation of insect population. pdf
- Type: Application
- Filed: February 25, 2015
- Publication date: August 27, 2015
- Inventors: Bruce A. Hay, Omar S. Akbari
(1) REPRESSIBLE LETHAL SYSTEM FOR REGULATING INSECT POPULATIONS
- Publication number: 20140283155
- Abstract: Disclosed herein are components, systems and methods for regulating population of multicellular organisms, for example insects. pdf
- Type: Application
- Filed: March 12, 2014
- Publication date: September 18, 2014
- Inventors: Omar S. Akbari, Katie L. Kennedy, Bruce A. Hay
