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Two RNAi-Related Patent Applications Published by the USPTO

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Title: Nucleic Acid and Corresponding Protein Entitled 98P4B6 Useful in Treatment and Detection of Cancer.

Number: 20050086707.

Filed: June 4, 2004.

Inventor: Aya Jakobovits, Agensys.

The patent application, its abstract states, cover "antibodies and molecules derived there from that bind to novel STEAP-1 protein, and variants thereof, … wherein STEAP-1 exhibits tissue specific expression in normal adult tissue, and is aberrantly expressed in [certain] cancers.

"Consequently, STEAP-1 provides a diagnostic, prognostic, prophylactic and/or therapeutic target for cancer," the abstract notes. "The STEAP-1 gene or fragment thereof, or its encoded protein, or variants thereof, or a fragment thereof, can be used to elicit a humoral or cellular immune response; antibodies or T cells reactive with STEAP-1 can be used in active or passive immunization."

The patent application specifically claims a STEAP-1 siRNA.


Title: Methods and Compositions for siRNA Expression.

Number: 20050089902.

Filed: Sept 1, 2004.

Lead Inventor: Lianxing Zheng, Scripps Research Institute.

The invention, the patent application's abstract states, comprises "one-step methods for generating siRNA expression cassettes. … Expression cassettes useful for siRNA are also provided."

Specifically, the application claims a "method for producing two complementary strands of a tripartite DNA comprising a 5' amplification primer binding site and a 3' amplification primer binding site with an intervening sequence between the two sites." This method involves "providing a polymerase extension reaction mixture comprising the following reagents: a) an initial double-stranded DNA comprising a left end comprising a 5' amplification primer binding site; and a right end complementary to a 3' end of an intervening linking primer; b) the intervening linking primer comprising: a 5' end complementary to a left end of a terminal DNA; a 3' end complementary to the right end of the initial DNA; and an intervening sequence between the 5' end and the 3' end; c) the terminal double-stranded DNA comprising a right end that is a 3' amplification primer binding site; and a left end complementary to the 5' end of the intervening linking primer; d) DNA polymerase and components sufficient for a polymerase chain reaction; reacting the reagents of the mixture in at least two thermocycles, wherein the first cycle comprises a melting temperature to denature double-stranded DNA; an initial annealing temperature to anneal the intervening linking primer to a strand of the initial double-stranded DNA; and an extension temperature for the polymerase to extend a 3' end of the strand of the initial double-stranded DNA using the intervening linking primer as a template to produce an intermediate product comprising the strand of the initial double-stranded DNA and a complement of the intervening linking primer; and the second cycle comprises a melting temperature to denature double-stranded DNA; an annealing temperature to anneal the intermediate product to a strand of the terminal double-stranded DNA; and an extension temperature for the polymerase to extend the 3' end of the intermediate product using the strand of the terminal double-stranded DNA as a template to produce a single-stranded tripartite DNA comprising the strand of the initial double-stranded DNA, the complement of the intervening linking primer, and the complement of the strand of the terminal double-stranded DNA sequences; and to extend the 3' end of the strand of the terminal double-stranded DNA using the intermediate product as a template to produce the complement of the single-stranded tripartite DNA."

The application further claims a kit for generating a siRNA polynucleotide, with "the kit comprising a) an initial double-stranded DNA comprising a left end comprising a 5' amplification primer binding site; and a right end complementary to a 3' end of an intervening linking primer; b) the intervening linking primer comprising: a 5' end complementary to a left end of a terminal DNA; and a 3' end complementary to the right end of the initial DNA; an intervening sequence between the 5' end and the 3' end; c) the terminal double-stranded DNA comprising a right end that is a 3' amplification primer binding site; and a left end complementary to the 5' end of the intervening linking primer; d) a 5' amplification primer; and e) a 3' amplification primer."

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