Eukaryotes expressing single stranded hybrid molecules
AssigneeThe University of Medecine and Dentistry of New Jersey
InventorsShohei Miyata, Atsushi Ohshima, Sumiko Inouye, Masayori Inouye
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Abstract
Eucaryotic cells like mammal and plant cells transfected with genes, called retrons, which code for stable single-stranded hybrid molecules (msDNA) containing RNA and DNA portions are disclosed. The retrons producing said linked RNA and DNA portions of the msDNAs also contain a gene encoding a reverse transcriptase (RT), which is necessary for the synthesis of the msDNAs. The msDNAs may contain foreign nucleic acid fragments which may be antisense fragments. These msDNA antisense vectors are useful for targeting genes of target proteins.
Claims30
A eucaryotic cell transfected with a DNA expression vector for replication, which vector contains a retron for msDNA synthesis, which retron contains msr and msd coding regions of the msDNA and a gene encoding a reverse transcriptase.
An expression vector which contains a retron for msDNA synthesis, which retron contains msr and msd coding regions of the msDNA and a gene encoding a reverse transcriptase.
An msDNA hybrid molecule which comprises a branched single-stranded RNA portion covalently linked to a single-stranded DNA portion by a 2',5'-phosphodiester bond between the 2'-OH group of an internal rG residue and the 5'-phosphate of the DNA molecule, which RNA is non-covalently linked to the DNA by base-pairing between the complementary 3' ends of the RNA and DNA molecules, which msDNA contains a stable stem-loop structure in the RNA and/or the DNA portion of the molecule, and which msDNA contains a foreign nucleic acid fragment.
Dependencies are inferred from the claim text.