公开(公告)号：US20200340028A1

公开(公告)日：2020-10-29

申请号：US16857563

申请日：2020-04-24

申请人： The University of Massachusetts

发明人： Craig Martin ,  Elvan Cavac ,  Kithmie Harshana Malagodapathiranage ,  Shuo Sui ,  Sarah L. Perry ,  Yasaman Gholamalipour

IPC分类号： C12P19/34 ,  C12N9/12 ,  C12N15/113

摘要： Described herein are synthetic methods for producing sequence-specific RNA oligonucleotides that eliminate impurities produced in prior art methods. In one aspect, an end-protected capture DNA complementary to a portion of the product RNA is employed. In another aspect, the template DNA is covalently or noncovalently linked to the RNA polymerase, either directly or through the use of a nontemplate DNA. In a third aspect, a flow chamber is employed. All of the methods can be used in combination.

公开(公告)号：US11578348B2

公开(公告)日：2023-02-14

申请号：US16857563

申请日：2020-04-24

申请人： The University of Massachusetts

发明人： Craig Martin ,  Elvan Cavac ,  Kithmie Harshana Malagodapathiranage ,  Shuo Sui ,  Sarah L. Perry ,  Yasaman Gholamalipour

IPC分类号： C12P19/34 ,  C12N9/12 ,  C12N15/10 ,  C12Q1/6865

摘要： Described herein are synthetic methods for producing sequence-specific RNA oligonucleotides that eliminate impurities produced in prior art methods. In one aspect, an end-protected capture DNA complementary to a portion of the product RNA is employed. In another aspect, the template DNA is covalently or noncovalently linked to the RNA polymerase, either directly or through the use of a nontemplate DNA. In a third aspect, a flow chamber is employed. All of the methods can be used in combination.

公开(公告)号：US20220112546A1

公开(公告)日：2022-04-14

申请号：US17512975

申请日：2021-10-28

申请人： The University of Massachusetts

发明人： Craig Martin ,  Elvan Cavac ,  Kithmie Harshana Malagodapathiranage

IPC分类号： C12Q1/6848 ,  C12Q1/6853

摘要： Described herein are synthetic methods for producing sequence-specific RNA oligonucleotides that eliminate impurities produced in prior art methods. In one aspect, a first amplification primer includes one or more deoxyuridine residues, wherein at least one of the one or more deoxyuridine residues is at position −1, −2, −3, −4 or −5 of the promoter region for the single-subunit, DNA-dependent RNA polymerase. The deoxyuridines are excised to provide an amplified functional template DNA which is then used to synthesize RNA which has reduced immunogenic double stranded RNA compared to controls.