Nucleic acid to activate gene expression and protein production
A protein and protein expression technology, applied in gene therapy, genetic material components, introduction of foreign genetic material using vectors, etc., can solve problems such as lack of sequences
Pending Publication Date: 2021-06-25
INST DE BIOLOGIA MOLECULAR E CELULAR IBMC
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However, the prior art still lacks sequences with all these features
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[0029]The inventors have discovered and characterized a novel short non-coding 28 nucleotide sequence, referred to herein as iPLUS - Protein Level Universal Sequence Increase - and specified in SEQ ID nr.1, which strongly enhances Protein production in vivo and in vitro.
[0030] Since this sequence is short, it can be obtained by chemical synthesis and can be easily inserted into an expression vector or the genome of a host cell. Surprisingly, it demonstrates a "universal" protein expression enhancer effect that is effective in multiple types of host cells and for different types of proteins of interest. Furthermore, because it is non-coding, its effects are achieved without interfering with the coding regions of genes and the proteins produced.
[0031] The iPLUS sequence (SEQ ID nr.1) can be obtained by nucleotide synthesis using solid-phase phosphoramidite nucleosides (as described in the following publications: Beaucage, SL; Iyer, RP (1992). amide method to synthesize o...
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The present invention refers to a method for increasing the efficiency of protein production. The referred method employs a composition that comprises a nucleic acid with a specific non- coding nucleotide sequence, for increasing protein expression. Another aspect of this invention refers the nucleic acid used in performing the aforementioned method, with a sequence identified as SEQ ID nr.1, or a truncated sequence of SEQ ID nr. 1, which strongly increases protein production in several eukaryotic systems, as well as the compositions that contain this nucleic acid sequence. The present invention also refers to genetic constructs or gene expression vectors comprising the nucleic acid with SEQ ID nr.1, as well as to the host cell lines comprising the genetic constructs or vectors incorporating the nucleic acid with SEQ ID nr.1. The present invention further refers to a kit comprising the above mentioned nucleotide sequence, compositions, vectors or the host cell lines for increasing protein expression, by conducting the present invention's method. The method, nucleic acid sequence, compositions, vectors, host cell lines and kit of the present invention can be advantageously used as a new tool to boost protein production in biotechnology and / or biopharmaceutical applications, due to its ability to enhance protein production derived from a genetic construct, a gene expression or a reporter vector, in zebra fish, fruit fly or other model organisms, as well as in mammalian or human cells. Thus, the present invention falls within the technical field of chemistry, biochemistry, molecular biology, microorganism's genetic engineering and recombinant-DNA technology.
Description
technical field [0001] The present invention relates to methods for increasing the efficiency of recombinant protein production by using compositions comprising nucleic acids having specific nucleotide sequences for gene incorporation in zebrafish, Drosophila or other model organisms, as well as mammalian or human cells constructs or sequences of gene expression vectors to enhance protein production and are advantageously used as new tools for increasing protein yields in biotechnological and / or biopharmaceutical applications. Therefore, the present invention belongs to the technical fields of chemistry, biochemistry, molecular biology, genetic engineering of microorganisms, and recombinant DNA technology. Background technique [0002] The efficiency of recombinant protein production is a significant bottleneck in most biopharmaceutical and biotechnology industries, and strategies to increase productivity often rely on improvements in host cell growth medium composition (as ...
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IPC IPC(8): C12N15/63C12N15/67
CPCC12N15/63C12N15/67A61K48/0066
Inventor M·奥利维拉A·杰西J·弗雷塔斯A·莫雷拉J·C·里贝罗贝萨
Owner INST DE BIOLOGIA MOLECULAR E CELULAR IBMC