130 results about "Terminal Repeats" patented technology

A method for producing recombinant adeno-associated virus in the absence of contaminating helper virus or wild-type virus involves culturing a mammalian host cell containing a transgene flanked by adeno-associated virus (AAV) inverse terminal repeats and under the control of regulatory sequences directing expression thereof, an AAV rep sequence and an AAV cap sequence under the control of regulatory sequences directing expression thereof, and the minimum adenovirus DNA required to express an E1a gene product, an E1b gene product and an E2a gene product, and isolating therefrom a recombinant AAV which expresses the transgene in the absence of contaminating helper virus or wildtype AAV. This method obviates a subsequent purification step to purify rAAV from contaminating virus. Also provided are various embodiments of the host cell.

Non-infectious, retrovirus-like particles contain mutations to reduce gag-dependent RNA-packaging of the gag gene product, eliminate reverse transcriptase activity of the pol gene product, eliminate integrase activity of the pol gene product and eliminate RNase H activity of the pol gene product through genetic manipulation of the gag and pol genes. The corresponding nucleic acid molecules are described. The non-infectious, retrovirus-like particles have utility in diagnosis.

A method of inactivating a proviral DNA integrated into the genome of a host cell latently infected with a retrovirus by treating the host cell with a composition comprising a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease, and two or more different guide RNAs (gRNAs), wherein each of the at least two gRNAs is complementary to a different target nucleic acid sequence in a long terminal repeat (LTR) in the proviral DNA, and inactivating the proviral DNA. A composition for use in inactivating a proviral DNA integrated into the genome of a host cell latently infected with a retrovirus including isolated nucleic acid sequences comprising a CRISPR-associated endonuclease and a guide RNA, wherein the guide RNA is complementary to a target sequence in a human immunodeficiency virus.

Some aspect of this invention provide nucleic acid constructs for transgene expression. Some aspects of this invention provide multicistronic nucleic acid constructs, for example, comprising an expression cassette encoding a hairpin RNA and a reporter expression cassette. Some aspects of this invention provide nucleic acid constructs comprising two or more self-complementary nucleic acid sequences, for example, hairpin RNA encoding nucleic acid sequences and AAV inverse terminal repeats. Methods for the use of the constructs in therapy and research are also provided.

This invention relates to modified parvovirus inverted terminal repeats (ITRs) that do not functionally interact with wild-type large Rep proteins, synthetic Rep proteins that functionally interact with the modified ITRs, and methods of using the same for delivery of nucleic acids to a cell or a subject. The modifications provide a novel Rep-ITR interaction that limits vector mobilization, increasing the safety of viral vectors.

This invention provides transcriptionally-activated AAV ITRs (inverted terminal repeats) which are small and transcriptionally active and uses thereof to optimize the expression of relatively large transgenes packaged in recombinant AAV vectors.

Non-infectious, non-replicating immunogenic HIV-like particles are produced by stable longn-term constitutive expression in mammalian cells by eliminating elements toxic to the mammalian cells. An expression vector contains a nucleic acid molecule comprising a modified HIV genome devoid of long terminal repeats and wherein Tat and vpr sequences are functionally disabled and a constitutive promoter operatively connected to the modified HIV genome for constitutive expression of the modified genome to produce the HIV-like particles.

In a wireless network structure which includes a wireless LAN device having a base station function (host device) which is compliant with IEEE802.11h and at least one wireless LAN device having a terminal function (slave device) which is not compliant with IEEE802.11h, the incompliant terminal transmits a probe request to the base station for confirming the presence of the base station. When the base station has dynamically switched its channel (frequency), a probe response is not returned from the base station to the terminal. Thus, the incompliant terminal transmits a probe request through a different channel in order to confirm the presence of the base station. The incompliant terminal repeats the above channel search process till the base station is found.

A method for quantification of an adeno-associated virus genome, including the steps of (a) preparing a composition containing a sample, at least one primer pair for use in amplification of only a nucleotide sequence contained in inverted terminal repeats of an adeno-associated virus, and an intercalating dye; (b) performing nucleic acid amplification reaction using the composition prepared in the step (a); and (c) detecting an amplified product obtained in the step (b). The present invention is especially useful in the fields of medicine, gene engineering, and biology.

An information communicating method provides a feedback mechanism, which transmits notice information to a client through a classified control server terminal. The notice information is transmitted to the client by the notice type of a preset lowest PRI and a client contact way corresponding to the notice type; simultaneously, a treatment state report of the notice information from a user is informed to a server terminal in real time by the feedback mechanism; if the client feeds back the treated information to the server terminal, the server terminal stops the information transmitting process; if not, the server terminal retransmits the notice information to the client by the notice type; if the client has no response in the preset time, the server terminal repeats the steps and informs the information report to the client by the notice type, the preset PRI of which is one grade higher than the notice type and repeats the steps until the server terminal receives the treated information fed back by the client or the PRI of the notice type reaches the highest preset level, thus improving the notice standard-reaching rate of the server terminal.

There are provided a method of communication which can reliably transmit information from a base station to a wireless terminal in a system which can reduce power consumption by intermittent operation in which the wireless terminal repeats operating state and suspended state by power on and off and the base station used for the method. The method includes the steps of storing base station information such as a command or data supplied to the wireless terminal in the base station, transferring the wireless terminal from the suspended state to the operating state to transmit information from the sensor to the base station, coupling the base station information stored in the base station to a response signal to transmit it to the wireless terminal in the operating state for transmitting the sensor information, and returning the wireless terminal to the suspended state after completing the transmission of the base station information and sensor information.

A complete virus-mediated system for recombinant AAV production including: (a) a first recombinant adenovirus encoding one or more AAV REP78/68 polypeptides and one or more viral capsid polypeptides; (b) a second recombinant adenovirus encoding a gene of interest and including AAV inverted terminal repeats, wherein the AAV inverted terminal repeats flank the gene of interest; (c) viral helper functions; and (d) a host cell transduced with the first recombinant adenovirus, the second recombinant adenovirus, and the viral helper functions.

A method of expressing in vivo heart-specific fluorescence in transgenic line of zebrafish is developed, which provides a research model for studying heart-related gene functions and performing gene therapies in the future. The method comprises the following steps. Firstly, a plasmid is constructed. This plasmid construct includes the upstream regulatory region, the exon 1, the intron 1, and the exon 2 of cmlc2 gene, cDNA of GFP, wherein the cmlc2 gene and GFP cDNA form a cassette, and inverted terminal repeats from adeno-associated virus are flanked at both sides of this cassette. The plasmid construct is linearized and microinjected into one-celled zebrafish fertilized eggs. Lastly, the heart-specific fluorescent expressed zebrafish are selected and the germline-transmitting transgenic strain is generated.

The instant invention provides silencer expression cassettes for delivery of RNAi therapy. The silencer cassettes of the instant invention encode an RNAi agent and lack means for replication within a host cell, inverted terminal repeats and further lacking one or more selectable markers. Optionally, the silencer cassettes lack CpG dinucleotides and/or the RNAi agent is within a scaffold of a naturally occurring miRNA. The composition comprising the silencer cassettes of the instant invention, as well as methods of use and suitable systems are also provided.

A helper dependent adenoviral vector system is provided. The subject helper dependent adenoviral vector system is made up of: (1) a "gutless" adenoviral vector, which in certain embodiments includes cis-acting human stuffer DNA that provides for in vivo long term, high level expression of a coding sequence present on the vector, where in certain embodiments the vector includes an integrating domain; (2) an adenoviral helper vector that is characterized by having an adenoviral genome region flanked by recombinase recognition sites, where the helper vectors further include a non-mammalian endonuclease recognition site positioned outside of the adenoviral genome region and in certain embodiments a third adenoviral inverted terminal repeat (ITR) sequence positioned between first and second terminal ITRs; and (3) a mammalian cell that expresses the corresponding recombinase and endonuclease, as well as the adenoviral preterminal and polymerase proteins. Also provided are methods of using the subject systems to produce virions having the subject helper dependent adenoviral vectors encapsulated in an adenoviral capsid. In addition, kits for use in practicing the subject methods are provided.