Isolation of hammerhead ribozymes with altered core sequences by in vitro selection

Biochemistry
N K VaishF Eckstein

Abstract

The hammerhead ribozyme has an invariant nucleotide sequence in the core region. In order to search for alternative sequences which can support the cleavage after the triplet GUC, the core region of 10 nucleotides was randomized and subjected to in vitro selection by repeated cycles of transcription, reverse transcription, and PCR. Active sequences were isolated after each transcription by denaturing PAGE, and after nine cycles of selection, two sequences dominated the pool. Both sequences conformed broadly to the consensus core region except that in one sequence a single A9U mutation was observed while in the other two mutations at A9U and U7A were seen. The catalytic efficiencies of these ribozymes were 6.4 and 14.1 microM(-1) min(-1), respectively, as compared to 163 microM(-1) min(-1) for the consensus sequence. Interestingly, the consensus was not found in any of the selected sequences. This discrimination against the consensus sequence was attributed to the specificity of the enzymes used in the selection procedure.

References

Jan 1, 1992·Annual Review of Biochemistry·R H Symons
Jun 25, 1992·Nucleic Acids Research·K J HertelR Hodgson

❮ Previous
Next ❯

Citations

Feb 13, 2001·Biopolymers·F Eckstein, B Bramlage
Nov 20, 1998·Current Opinion in Biotechnology·P J WelchF Wong-Staal
Sep 8, 2000·Bioorganic & Medicinal Chemistry·A R KoreF Eckstein
May 10, 2000·Chemistry & Biology·A BeaudryL Beigelman
Jul 17, 1998·Current Opinion in Chemical Biology·S VermaF Eckstein
Aug 6, 1998·Current Opinion in Chemical Biology·M Famulok, A Jenne
Mar 29, 2000·Nucleic Acids Research·M P Robertson, A D Ellington
Oct 6, 1998·Annual Review of Biochemistry·S Verma, F Eckstein
Apr 16, 1998·Proceedings of the National Academy of Sciences of the United States of America·N K VaishF Eckstein
May 24, 2000·Proceedings of the National Academy of Sciences of the United States of America·J Tang, R R Breaker
Aug 20, 2011·Wiley Interdisciplinary Reviews. RNA·Katarzyna Matylla-KulinskaRenée Schroeder
Mar 26, 2009·Chemistry & Biology·Maung Nyan WinChristina D Smolke
May 19, 2017·Organic & Biomolecular Chemistry·Getong LiuZhuo Tang
Jan 17, 2019·Nucleic Acids Research·Xin HuangXianming Mo
Jan 14, 2016·Interdisciplinary Sciences, Computational Life Sciences·Angela Gupta, D Swati

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Proceedings of the National Academy of Sciences of the United States of America
N K VaishF Eckstein
Proceedings of the National Academy of Sciences of the United States of America
D M Long, O C Uhlenbeck
PCR Methods and Applications
P DeGrandisM Altschuler
© 2021 Meta ULC. All rights reserved