Unnatural base pairs for specific transcription.
An unnatural base pair of 2-amino-6-(N,N-dimethylamino)purine (designated as x) and pyridin-2-one (designated as y) has been developed for specific transcription. The ribonucleoside triphosphates of y and a modified y, 5-methylpyridin-2-one, are selectively incorporated into RNA opposite x in the templates by T7 RNA polymerase. In addition, the sequences of the DNA templates containing x can be confirmed by a dideoxynucleotide chain-terminator method supplemented with the deoxynucleoside triphosphate of y. The bulky dimethylamino group of x in the templates effectively eliminates noncognate pairing with the natural bases. These results enable RNA biosynthesis for the specific incorporation of unnatural nucleotides at the desired positions.
Ohtsuki
T
T
Genomic Sciences Center, Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology Corporation (JST), Institute of Physical and Chemical Research (RIKEN), 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan.
Kimoto
M
M
Ishikawa
M
M
Mitsui
T
T
Hirao
I
I
Yokoyama
S
S
D020029:Base Pairing / Q000235:genetics*; D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D008969:Molecular Sequence Data; D011687:Purines / Q000378:metabolism*; D011728:Pyridones / Q000378:metabolism*; D012313:RNA / Q000096:biosynthesis; D017422:Sequence Analysis, DNA; D013698:Templates, Genetic; D014158:Transcription, Genetic / Q000235:genetics*; D014764:Viral Proteins
C425930:2-amino-6-(N,N-dimethylamino)purine; D011687:Purines; D011728:Pyridones; D014764:Viral Proteins; D012313:RNA; D004247:DNA; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases