초록 ▼

The present invention is directed to collections of templates for molecules such as RNA, as well as templates, devices, kits and methods for generating molecules from these collections. Through the use of various embodiments of the present invention one may efficiently and effectively obtain selecte

The present invention is directed to collections of templates for molecules such as RNA, as well as templates, devices, kits and methods for generating molecules from these collections. Through the use of various embodiments of the present invention one may efficiently and effectively obtain selected RNA molecules such as siRNA, shRNA, miRNA mimics and inhibitors, lncRNA, antisense RNA, aptamers, ribozymes, and sgRNA and sets of those molecules.

대표청구항 ▼

1. A library of DNA molecules, wherein the library comprises a plurality of DNA molecules, wherein each DNA molecule comprises a sequence of interest located between a first primer binding region and a second primer binding region, wherein for each DNA molecule the combination of the first primer bi

1. A library of DNA molecules, wherein the library comprises a plurality of DNA molecules, wherein each DNA molecule comprises a sequence of interest located between a first primer binding region and a second primer binding region, wherein for each DNA molecule the combination of the first primer binding region and the second primer binding region is unique to the sequence of interest and within each DNA molecule the sequence of interest is located upstream of one of the first primer binding region and the second primer binding region, and downstream of the other of the first primer binding region and the second primer binding region, and within a first subset of DNA molecules, each of a plurality of DNA molecules has the same first primer binding region and a different second primer binding region and within a second subset of DNA molecules, each of a plurality of DNA molecules has the same first primer binding region and a different second primer binding region, wherein the first primer binding region of the first subset is distinct from the first primer binding region of the second subset and wherein if a sequence of interest codes for or is complementary to a molecule of an organism, less than 50% of the sequence of nucleotides immediately upstream and downstream of the sequence of interest is identical to or complementary to the sequence of nucleotides in the regions immediately upstream and downstream of the molecule of the organism. 2. The library of claim 1, wherein the library comprises at least 100 DNA molecules. 3. The library of claim 2, wherein there are at least 20 subsets, wherein each subset contains at least five DNA molecules, wherein in each subset each DNA molecule has the same first primer binding region and a different second primer binding region. 4. The library of claim 3, wherein, the first primer binding region is a reverse primer binding region and the second primer binding region is a forward primer binding region and each DNA molecule further comprises a first universal primer binding region and a second universal primer binding region, wherein for each DNA molecule, the forward primer binding region, the sequence of interest and the reverse primer binding region are located between the first universal primer binding region and the second universal primer binding region. 5. The library of claim 4, wherein each DNA molecule comprises a first restriction site and a second restriction site, wherein the first restriction site is located on one side of the sequence of interest and the second restriction site is located on the other side of the sequence of interest. 6. The library of claim 5, wherein the sequence of interest codes for a sequence selected from the group consisting of an siRNA, an shRNA, an miRNA mimic, an miRNA inhibitor, an lncRNA, an antisense RNA, an aptamer, a ribozyme and a small guide RNA, or codes for a complement of a sequence selected from the group consisting of an siRNA, an shRNA, an miRNA mimic, an miRNA inhibitor, an lncRNA, an antisense RNA, an aptamer, a ribozyme and a small guide RNA. 7. The library of claim 4, wherein each DNA molecule further comprises a T7 promoter sequence, wherein the T7 promoter sequence is located 5′ of the sequence of interest. 8. The library of claim 1, wherein each DNA molecule further comprises a T7 promoter sequence, wherein the T7 promoter sequence is located 5′ of the sequence of interest and the sequence of interest is an shRNA, wherein the shRNA comprises an antisense region, a loop region and a sense region. 9. The library of claim 1, wherein the first primer binding region is a forward primer binding region and the second primer binding region is a reverse primer binding region and the library further comprises a third subset, wherein each DNA molecule within the third subset has the same reverse primer binding region and a different forward primer binding region, and a fourth subset, wherein each DNA molecule within the fourth subset has the same reverse primer binding region and a different forward primer binding region and the reverse primer binding region of the fourth subset is distinct from the reverse primer binding region of the third subset. 10. A library of DNA molecules, wherein the library comprises a plurality of DNA molecules, wherein each DNA molecule comprises a sequence of interest located between a first primer binding region and a second primer binding region, wherein for a first group of sequences of interest, each sequence of interest is located between a first unique combination of the first primer binding region and the second primer binding region and for a second group of sequences of interest, each sequence of interest is located between a second unique combination of the first primer binding region and the second primer binding region, wherein within the library of DNA molecules the first group of sequences of interest comprises a plurality of sequences that code for a target of a first common pathway and the second group of sequences of interest comprises a plurality of sequences of interest that code for a target of a second common pathway, wherein within each DNA molecule the sequence of interest is located upstream of one of the first primer binding region and the second primer binding region, and downstream of the other of the first primer binding region and the second primer binding region, wherein the first combination is different from the second combination, and wherein if a sequence of interest codes for or is complementary to a molecule of an organism, less than 50% of the sequence of nucleotides immediately upstream and downstream of the sequence of interest is identical to or complementary to the nucleotides in the regions immediately upstream and downstream of the molecule of the organism. 11. The library of claim 10, wherein the first group of sequences of interest codes for a region of a first gene, and the second group of sequences of interest codes for a region of a second gene. 12. A method for isolating a group of nucleotide molecules of interest comprising: a. exposing the library of claim 4 to a first primer and a second primer, wherein either: (i) the first primer is capable of binding to the first universal primer binding region and the second primer is capable of binding to the reverse primer binding region of a group of DNA molecules, or(ii) the first primer is capable of binding to the second universal primer binding region and the second primer is capable of binding to the forward primer binding region of a group of DNA molecules;b. conducting PCR to generate a PCR product; andc. isolating a group of nucleotide molecules of interest from the PCR product. 13. A method of cloning a DNA sequence comprising: a. inserting a nucleotide of interest from the group of nucleotides of interest of claim 12 into an expression vector thereby generating an expression vector comprising the nucleotide molecule of interest;b. delivering the expression vector comprising the nucleotide molecule of interest to a cell; andc. exposing the cell to conditions that permit cloning to occur. 14. The method according to claim 13, wherein said delivering is via viral particles. 15. A method of cloning a group of DNA sequences comprising: a. inserting the group of nucleotide molecules of interest of claim 12 into expression vectors thereby generating expression vectors comprising the nucleotide molecules of interest;b. delivering the expression vectors comprising the nucleotide molecules of interest to cells; andc. exposing the cells to conditions that permit cloning to occur. 16. The method according to claim 15, wherein said delivering is via viral particles.