New Invention Profile
| Publication No: | IN202211075512 [India] | Application No: | 202211075512 |
| Title: | Engineering 5' transcript of human therapeutics for enhanced production in a heterologous expression system | ||
| Publication Date: | 28-06-2024 | File Date: | 26-12-2022 |
| Inventor(s): | Sonal Datta; Akshit Vikram Jain | ||
| IPC Classification: | C12N 15/67, C07K 16/28, C12P 21/02, C12N 15/63, C12N 15/70 | ||
| Abstract: | Differential preference in codon usage in each organism could impede translation due to the requirement for specific tRNAs that may be in low abundance or lacking in a surrogate host. Furthermore, the formation of complex secondary structures towards the 5' of the transcript could impede translation efficiency. The present disclosure relates to enhancing protein expression through 5' engineering of transcript sequence concerning mRNA secondary structure, GC content, rare codon replacement, and codon preference of the host without codon optimization of the entire gene sequence. The invention discloses that the mRNA secondary structure proximal to the start codon is the critical limiting factor for an efficient expression of G-CSF protein in E. coli. The invention also describes a simple and effective method for the high-level production of proteins by optimizing the 5' end of the transcript/gene sequence. Herein invention discloses through mRNA engineering of mere two codons succeeding initiation codon in codon-optimized G-CSF and native G-CSF sequence, results in up to seven folds and twenty-eight folds increase in protein expression, respectively. |
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Industrial Applications
Biological Science Processes/Methods [Biological Science]
Biomolecules [Biological Science]
Biotechnology [Biological Science]