Abstract: The present disclosure provides transgenic nematode systems for assessing function of heterologous genes, their variants and drug discovery. The transgenic nematodes contain a heterologous gene that is inserted via homologous recombination at the native locus replacing and removing the nematode ortholog, wherein expression of the heterologous gene rescues function of the removed nematode ortholog and a transgenic control animal is provided. The heterologous gene may be further modified to provide a variant, such as a human clinical variant, whereby a transgenic test animal is provided. Those transgenic test animals are used in methods to assess function of the heterologous variant and drug screens to find therapeutic candidates reversing deviant activity back to wildtype.

Abstract: This disclosure relates to methods for screening therapeutic agents to treat altered function of a mutated target gene (e.g., clinical variant) as well as reagents for use in the same.

Abstract: Disclosed herein are microinjection chips, devices, and systems for injection of unicellular or multicellular organisms. The microinjection chip and device disclosed herein include the microfluidic features, inlet port, pre-injection reservoir, injection channel and post injection channel in fluid communication with each other. The inlet port is adapted to sequentially move individual organisms into the injection channel, which is adapted to immobilize the individual organism in fluid. The injection channel features a side wall adapted to receive a microinjection pipette without a microinjection port and to reseal when the microinjection pipette is removed.

Abstract: The present disclosure provides transgenic non-human animal (e.g., nematode) systems for assessing heterologous polygenic or monogenic phenotypes, their variants and drug discovery. The transgenic non-human animals (e.g., nematodes) contain a first heterologous polypeptide coding sequence and a second heterologous polypeptide coding sequence (a plurality of heterologous polypeptide coding sequences), wherein the first and second heterologous polypeptide coding sequences are integrated into the host animal genome, and wherein expression of the first and second heterologous polypeptide coding sequence contribute to the heterologous phenotype. The plurality of heterologous polypeptide coding sequences are interrelated wherein their expression products, directly or indirectly, contribute or lead to an observable phenotype.

Abstract: The present disclosure provides a microfluidic device and system for measuring a composite electropharyngeogram (EPG) signal from a pool of multiple nematodes, wherein the composite EPG signal is measured from the pool of nematodes present in a single recording channel connected to two or more integrated electrodes. The microfluidic device includes an inlet port and outlet port directly connected to a single recording channel and two or more electrodes directly connected to the recording channel. The recording channel is configured to hold 10 to 10,000 nematodes.

Abstract: Disclosed herein are classifier models, computer implemented systems, machine learning systems and methods thereof for classifying clinical variants of unknown or uncertain significance into a pathogenicity category using measured phenotype features extracted from phenotype assays of transgenic organism expressing the human clinical variant. Embodiments of the present invention relate generally to methods for generating classifier models using machine learning and use of those classifier models to predict the pathogenicity of a clinical variant for a specific human disease (e.g. genetic disease), assigning a patient clinical variant to a pathogenicity category (e.g. pathogenic or benign) for the specific human disease to determine whether that patient should be followed up with additional, more invasive diagnostic testing, or treatment.

Abstract: The present disclosure provides transgenic nematode systems for assessing function of heterologous genes, their variants and drug discovery. The transgenic nematodes contain a heterologous gene that is inserted via homologous recombination at the native locus replacing and removing the nematode ortholog, wherein expression of the heterologous gene rescues function of the removed nematode ortholog and a transgenic control animal is provided. The heterologous gene may be further modified to provide a variant, such as a human clinical variant, whereby a transgenic test animal is provided. Those transgenic test animals are used in methods to assess function of the heterologous variant and drug screens to find therapeutic candidates reversing deviant activity back to wildtype.

Abstract: The present disclosure provides a microfluidic device and system for measuring a composite electropharyngeogram (EPG) signal from a pool of multiple nematodes, wherein the composite EPG signal is measured from the pool of nematodes present in a single recording channel connected to two or more integrated electrodes. The microfluidic device includes an inlet port and outlet port directly connected to a single recording channel and two or more electrodes directly connected to the recording channel. The recording channel is configured to hold 10 to 10,000 nematodes.

Abstract: The present disclosure provides homologous recombination reporter nucleic acid construct reagents for increasing the likelihood of detecting successful modification of a specific sequence in chromosomal DNA of a host cell via homologous recombination. The homologous recombination reporter constructs contain a sequence element inserted within the coding sequence for a reporter gene resulting in a mutated reporter gene. The sequence element is removed via homologous recombination based on the presence of two homology regions present in the reporter construct.

Abstract: Embodiments of the present invention provide materials and methods for performing phenotype-based chemical screens. Embodiments disclosed herein pertain to the use of a battery of high-throughput zebrafish behavioral assays to generate behavioral profiles. In accordance with these embodiments, responses to various chemical compounds with known and unknown biological functions can be used to generate behavioral profiles. Embodiments also involve generating behavioral profiles based on genetic mutations or environmental perturbations. Establishing databases of behavioral profiles facilitates identification of novel chemicals that phenocopy effects of therapeutic agents and/or that modulate genetic or environmental behavioral profiles, thereby providing a basis for not only assessing the properties of known chemical compounds but also for developing novel treatments for human diseases.

Abstract: The present disclosure provides transgenic nematode systems for assessing function of heterologous genes, their variants and drug discovery. The transgenic nematodes contain a heterologous gene that is inserted via homologous recombination at the native locus replacing and removing the nematode ortholog, wherein expression of the heterologous gene rescues function of the removed nematode ortholog and a transgenic control animal is provided. The heterologous gene may be further modified to provide a variant, such as a human clinical variant, whereby a transgenic test animal is provided. Those transgenic test animals are used in methods to assess function of the heterologous variant and drug screens to find therapeutic candidates reversing deviant activity back to wildtype.

Abstract: The present disclosure provides methods and systems for performing, observing, and/or recording defecation motor program (DMP) events using microfluidic devices. The methods may be performed wherein the nematodes ingest fluorescent or color material and are then loaded in a microfluidic chip and stimulated to feed and defecate. DMP events are observed with use of a fluorescent microscope. In other methods, a microfluidic device with two or more electrodes is used to record electrical events of the DMP.