A New Method for Synthesizing RNA on Silica Supports
Until recently we perceived that there was only a minimal need for methods for chemical synthesis of RNA. This was because almost any RNA sequence could be synthesized using either the SP6 or T7 promoter and an appropriate DNA duplex (1,2). However, several recent developments primarily centered on studies of intron splicing mechanisms (3–6) have led us to initiate a program in this area. We now feel that eventually new chemical methods for synthesizing RNA will be very important as an aid to helping us understand a large number of biochemical processes such as mRNA and tRNA maturation (3–6), conformational analysis of RNA (7), and the mechanisms whereby proteins recognize and bind to RNAs (8). For example, we will probably want to construct mRNA, lariat and branched tetranucleotide structures, tRNAs, viroids, 5S RNA or even ribosomal RNA in such a way that sugar or base analogues are inserted at certain key sites so that their biochemical activity can be studied. We would envisage such a synthesis to involve first preparing the analogue portion of the RNA chemically and the remainder from DNA and either the T7 or SP6 promoter. These segments would then be joined with T4-RNA ligase (1) to form the RNA of interest.
KeywordsThin Layer Chromatography Flash Chromatography Calcium Hydride Phosphinic Acid Thin Layer Chromato
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