Biotechnological Applications of Protein Splicing

Abstract

Protein splicing domains, also called inteins, have become a powerful biotechnological tool for applications involving molecular biology and protein engineering. Early applications of inteins focused on self-cleaving affinity tags, generation of recombinant polypeptide α-thioesters for the production of semisynthetic proteins and backbone cyclized polypeptides. The discovery of naturallyoccurring split-inteins has allowed the development of novel approaches for the selective modification of proteins both in vitro and in vivo. This review gives a general introduction to protein splicing with a focus on their role in expanding the applications of intein-based technologies in protein engineering and chemical biology.

Suprachoroidal Delivery of Viral and Nonviral Gene Therapy for Retinal Diseases

“…Recent AAV technological advances are under preclinical investigation and are being considered for clinical development. This technology includes dual AAV 53–55 and triple AAV, 56 in which case, a large transgene is split into 2 or 3 separate vectors to ultimately reconstitute to a full-length gene, as well as intein-mediated protein trans-splicing, 57 which reconstitute large proteins from shorter precursor polypeptides (separately encoded by 2 or more independent AAV vectors). Although convincible preclinical data have been generated through subretinal administration, 58 these platforms have not yet been assessed by the SC route.…”

Preparing Proteoforms of Therapeutic Proteins for Top-Down Mass Spectrometry

“…The heterogeneity is developing during the production of an rTP mainly in the upstream processing. The first event increasing the heterogeneity is alternative splicing [17][18][19]. The second critical step is the protein biosynthesis at the ribosomes, in which errors can occur.…”

Off‐Pathway‐Sensitive Protein‐Splicing Screening Based on a Toxin/Antitoxin System

“…The unique autocatalytic reaction catalyzed by inteins, which is capable of ligating protein domains, has enabled diverse biotechnological applications of inteins such as protein purification, ligation, and chemical modification . Protein splicing can not only occur in cis but also in trans , through artificially or naturally split inteins, leading to the ligation of individual polypeptides.…”