Suppressor tRNAs at the interface of genetic code expansion and medicine

Awawdeh, Aya; Radecki, Alexander A.; Vargas-Rodriguez, Oscar

doi:10.3389/fgene.2024.1420331

Cited by 2 publications

(1 citation statement)

References 78 publications

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“…The aberrant generation of a stop codon in a gene is the cause of nearly 11% of all human genetic disorders ( Mort et al, 2008 ). As such, recent research has sought to develop safe and efficient suppressor tRNAs to readthrough pathogenic stop codons and rescue disrupted mRNAs and truncated proteins ( Porter et al, 2021 ; Awawdeh et al, 2024 ; Coller and Ignatova, 2024 ). tRNA suppression therapy is at an early stage of development, with most research at the pre-clinical phase.…”

Section: Therapeutic Applications Of Trna Engineeringmentioning

confidence: 99%

Tuning tRNAs for improved translation

Weiss,

Decker,

Bolano

et al. 2024

Front. Genet.

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Transfer RNAs have been extensively explored as the molecules that translate the genetic code into proteins. At this interface of genetics and biochemistry, tRNAs direct the efficiency of every major step of translation by interacting with a multitude of binding partners. However, due to the variability of tRNA sequences and the abundance of diverse post-transcriptional modifications, a guidebook linking tRNA sequences to specific translational outcomes has yet to be elucidated. Here, we review substantial efforts that have collectively uncovered tRNA engineering principles that can be used as a guide for the tuning of translation fidelity. These principles have allowed for the development of basic research, expansion of the genetic code with non-canonical amino acids, and tRNA therapeutics.

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Section: Therapeutic Applications Of Trna Engineeringmentioning

confidence: 99%

Tuning tRNAs for improved translation

Weiss,

Decker,

Bolano

et al. 2024

Front. Genet.

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Efficient suppression of premature termination codons with alanine by engineered chimeric pyrrolysine tRNAs

Awawdeh,

Tapia,

Alshawi

et al. 2024

Nucleic Acids Research

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Mutations that introduce premature termination codons (PTCs) within protein-coding genes are associated with incurable and severe genetic diseases. Many PTC-associated disorders are life-threatening and have no approved medical treatment options. Suppressor transfer RNAs (sup-tRNAs) with the capacity to translate PTCs represent a promising therapeutic strategy to treat these conditions; however, developing novel sup-tRNAs with high efficiency and specificity often requires extensive engineering and screening. Moreover, these efforts are not always successful at producing more efficient sup-tRNAs. Here we show that a pyrrolysine (Pyl) tRNA (tRNAPyl), which naturally translates the UAG stop codon, offers a favorable scaffold for developing sup-tRNAs that restore protein synthesis from PTC-containing genes. We created a series of rationally designed Pyl tRNAScaffold Suppressor-tRNAs (PASS-tRNAs) that are substrates of bacterial and human alanyl-tRNA synthetase. Using a PTC-containing fluorescent reporter gene, PASS-tRNAs restore protein synthesis to wild-type levels in bacterial cells. In human cells, PASS-tRNAs display robust and consistent PTC suppression in multiple reporter genes, including pathogenic mutations in the tumor suppressor gene BRCA1 associated with breast and ovarian cancer. Moreover, PTC suppression occurred with high codon specificity and no observed cellular dysregulation. Collectively, these results unveil a new class of sup-tRNAs with encouraging potential for tRNA-based therapeutic applications.

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Suppressor tRNAs at the interface of genetic code expansion and medicine

Cited by 2 publications

References 78 publications

Tuning tRNAs for improved translation

Tuning tRNAs for improved translation

Efficient suppression of premature termination codons with alanine by engineered chimeric pyrrolysine tRNAs

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