(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
