The present invention provides robust, streamlined, reproducible and highly efficient eukaryotic cell transfection systems and related methods. The highly-efficient systems and methods of the present invention reduce the number of steps required to transfect cells and reduce, e.g., eliminate, the need for specialized equipment. In particular, the systems and related methods afford the ability for streamlining transfection while retaining and improving robust and reproducible transfection efficiencies, cell viability, and/or protein production. Furthermore, the highly-efficient systems and methods of the present invention for transfecting eukaryotic cells also eliminate the need for any specialized or complicated preparation of exogenous nucleic acid, which makes available high throughput and/or large scale transfection.

The present invention relates to the production of adeno-associated viral vectors in insect cells. The insect cells therefore comprise a first nucleotide sequence encoding the adeno-associated virus (AAV) capsid proteins, whereby the initiation codon for translation of the AAV VP1 capsid protein is an AUG. Upstream of the VP1 open reading frame an alternative out of frame start codon is placed such that translation initiation of the VP1 protein is modified, i.e. reduced, to allow production of VP1:VP2:VP3 in a good stoichiometry resulting in AAV with high potency.

Methods and compositions relating to the engineering of an improved protein with methionine--lyase enzyme activity are described. For example, in certain aspects there may be disclosed a modified cystathionine--lyase (CGL) comprising one or more amino acid substitutions and capable of degrading methionine. Furthermore, certain aspects of the invention provide compositions and methods for the treatment of cancer with methionine depletion using the disclosed proteins or nucleic acids.

The present invention provides a primary cell culture which combines a cell culture medium and cells derived from a hypertrophied androgenic gland (AG) of a decapod crustacean. The invention also provides methods for obtaining an all-female progeny by initially injecting/transplanting the primary cell culture to a genetic-female to obtain a male-Neo-male.

By establishing effective methods for shrimp 3D cell culture and passage, the present invention provides a technology of continuous shrimp cell culture intended for the establishment of immortalized shrimp cell lines. The present invention provides a preparation method of matrigel for 3D cell culture of shrimp by optimizing an additive proportion of matrigel. The present invention further provides a technology of separation and 3D cell culture of shrimp haemolymph cells, where shrimp haemolymph cells adhere to and grow on the surface of the matrigel in the form of a single round cell and a cell pellet/cellular spheroid, with survival and growth abilities being superior to 2D culture effects. The above technology is achieved by optimizing a formula of complete medium for shrimp cells, selecting the medium as an anticoagulant and a diluent for shrimp haemolymph cells, selecting a 3D culture method for surface-adhered growth in the matrigel.

The present invention relates to means and methods for increasing the efficiency of recombinant protein expression and, more particularly, to means and methods for optimizing the industrial production of recombinant proteins in pyralid moths eggs (which are insect pests of stored foods), especially Mediterranean flour moth eggs (Ephestia kuehniella). Furthermore, the present invention relates to the pyralid moths eggs itself which contains a recombinant baculovirus and, infection of recombinant baculovirus into pyralid moths eggs and, transformation or transduction or transfection by recombinant baculoviruses or bacmids. In addition, it relates to an appropriate device for carrying out the method of the present invention.

Provided herein are methods and compositions useful in the production of recombinant AAV (rAAV) in host producer cells, such as insect cells. In some embodiments, methods, uses and compositions are provided that comprise recombinant VP1 genes comprising modified Kozak sequences to express AAV VP1 proteins in amounts that are useful for producing infective rAAV particles. These infective rAAV particles may comprise a gene of interest.

The present invention provides a primary cell culture which combines a cell culture medium and cells derived from a hypertrophied androgenic gland (AG) of a decapod crustacean. The invention also provides methods for obtaining an all-female progeny by initially injecting/transplanting the primary cell culture to a genetic-female to obtain a male-Neo-male.

The present invention is directed to recombinantly-modified adeno-associated virus (AAV) helper vectors that are capable of increasing the packaging efficiency of recombinantly-modified adeno-associated virus (rAAV) and their use to improve the packaging efficiency of such rAAV. The present invention is particularly directed to recombinantly-modified adeno-associated virus (AAV) helper vectors that have been further modified to replace (or augment) the P5 and/or P40 promoter sequences that are natively associated with the Rep proteins encoded by such rAAV with AAV P5 and/or P40 promoters that are associated with the Rep proteins of an rAAV of different serotype. The use of such substitute or additional promoter sequences causes increased production of recombinantly-modified adeno-associated virus.

An insect culture maintenance system includes a sequence of open-ended cylindrical tubes [500, 502, 504, 506] joined pairwise alternately at their tops and bottoms using multiple dual-cap connectors. Each dual-capped connector has a channel from an inside of a first cap to an inside of a second cap. In use, connectors that cap the bottoms of the tubes [508, 512] are filled with insect food media [518, 520], while connectors that cap the tops of the tubes [510] are open. As a result of this design, adults pass from one tube to the next through the top dual-cap connectors, while larvae pass from one tube to the next through the bottom dual-cap connectors, resulting in propagation of subsequent generations of insects through the sequence of tubes.