Kithmie MalagodaPathiranage, Ruptanu Banerjee, and Craig T. Martin, Nucleic Acids Research, 52(17), 10607-10618. 2024 Current approaches to RNA synthesis/manufacturing require substantial (and incomplete) purification post-synthesis. We have previously demonstrated the synthesis of RNA from a complex in which T7 RNA polymerase …

A New Approach to RNA Synthesis: Immobilization of Stably and Functionally Co-Tethered Promoter DNA and T7 RNA Polymerase Read more »

Kithmie MalagodaPathiranage and Craig T. Martin, Methods in Enzymology, 691, 209-222, 2023 T7 RNA polymerase is widely used to synthesize RNA of any length, and long-standing protocols exist to efficiently generate large amounts of RNA. Such synthesis, however, is often plagued …

A Simple Approach to Improving RNA Synthesis: Salt Inhibition of RNA Rebinding Coupled With Strengthening Promoter Binding by a Targeted Gap in the DNA Read more »

Kithmie MalagodaPathiranage, Elvan Cavac, Tien-Hao Chen, Bijoyita Roy, Craig T Martin Nucleic Acids Research, 51(6), e36 (12 pages) T7 RNA polymerase is commonly used to synthesize large quantities of RNA for a wide variety of applications, from basic science to …

High-salt transcription from enzymatically gapped promoters nets higher yields and purity of transcribed RNAs Read more »

Yasaman Gholamalipour, William C. Johnson, & Craig T. Martin, Nucl Acids Res, 47, e118 (7 pages). 2019 In vitro synthesized RNA is used widely in studies of RNA biology, biotechnology and RNA therapeutics. However, in vitro synthesized RNA often contains impurities, such as …

Efficient inhibition of RNA self-primed extension by addition of competing 3’-capture DNA – improved RNA synthesis by T7 RNA polymerase Read more »

Satamita Samanta & Craig T. Martin, J. Biol. Chem. 288, 31993-32003, 2013. It has long been known that during initial transcription of the first 8-10 bases of RNA, complexes are relatively unstable, leading to the release of short abortive RNA transcripts. An …

Insights into the Mechanism of Initial Transcription in E coli RNA Polymerase. Energetic Mechanisms Leading to Initial Complex Instability and Abortive Cycling Read more »

Luis Ramírez-Tapia & Craig T. Martin, J. Biol. Chem. 287, 37352-37361, 2012. RNA polymerases undergo substantial structural and functional changes in transitioning from sequence-specific initial transcription to stable and relatively sequence-independent elongation. Initially transcribing complexes are characteristically unstable, yielding short abortive products …

New Insights into the Mechanism of Initial Transcription: the T7 RNA Polymerase Mutant P266L Transitions to Elongation at Longer RNA Lengths than Wild Type Read more »

Ankit V. Vahia & Craig T. Martin, Biochemistry 50, 7015-7022, 2011.  Although the synthesis of RNA from a DNA template is (and must be) a generally very stable process to enable transcription of kilobase transcripts, it has long been known that …

Direct Tests of the Energetic Basis of Abortive Cycling in Transcription Read more »

Xiaoqing Liu & Craig T. Martin, J. Biol. Chem. 284, 36262-36270, 2009. Transcription machinery from a variety of organisms shows striking mechanistic similarity. Both multi- and single subunit RNA polymerases have evolved an 8-10 base pair RNA:DNA hybrid as a part of …

Transcription Elongation Complex Stability: The Topological Lock Read more »