Reverse Transcriptase: From Transcriptomics to Genome Editing

Area of Science:

• Biochemistry
• Molecular Biology
• Genomics

Background:

• Reverse transcriptases (RTs) synthesize complementary DNA (cDNA) from RNA templates.
• RTs coupled with PCR are standard for RNA detection and gene cloning.
• Protein engineering has significantly improved classical retroviral RTs.

Purpose of the Study:

• To review the properties and biotechnological applications of wild-type and engineered RTs.
• To highlight the role of RTs in advancing transcriptomics and next-generation sequencing.
• To discuss the emerging use of engineered RTs in genome manipulation.

Main Methods:

• Review of existing literature on reverse transcriptase enzymes.
• Analysis of protein engineering strategies for RT improvement.
• Examination of RT applications in molecular biology techniques.

Main Results:

• Engineered RTs are central to next-generation sequencing and transcriptomics.
• RTs fused with CRISPR/Cas9 nickase show promise for eukaryotic genome editing.
• New RTs and engineered variants expand biotechnological capabilities.

Conclusions:

• Reverse transcriptases are versatile tools with expanding applications.
• Protein engineering continues to enhance RT functionality for diverse uses.
• RTs are integral to modern molecular biology, from diagnostics to genome editing.