- Ribonuclease T1 (RNase T1) is a guanosine-specific endoribonuclease that cleaves single-stranded RNA at the 3′-end of guanine residues, generating RNA fragments with 3′-phosphate and 5′-hydroxyl termini. It is a microbial enzyme, most commonly isolated from the fungus Aspergillus oryzae. RNase T1 is highly sequence-specific, making it a valuable tool in RNA sequencing and structural analysis.
- Unlike general RNases such as RNase A, RNase T1 does not cleave at other nucleotides, providing precise cleavage patterns that are instrumental in RNA fingerprinting, oligonucleotide mapping, and secondary structure studies. It requires mild conditions for activity, typically in the presence of low concentrations of salts and a near-neutral pH. RNase T1 is most effective under denaturing conditions (e.g., with urea), which prevent secondary structure formation and allow complete cleavage at all guanine residues.
- RNase T1 is widely used in the analysis of RNA modifications, mutational studies, and RNA folding. It is often used in combination with other RNases, such as RNase A or RNase V1, to generate complementary digestion patterns. The enzyme is also employed in the quality control of synthetic RNA and in the production of RNA ladders for gel electrophoresis.
- In summary, RNase T1 is a sequence-specific endoribonuclease that cleaves RNA at guanine residues. Its precision and reliability make it an essential tool in RNA structure and sequence analysis, offering insights into RNA processing, folding, and interactions.
