Protein Splicing of Inteins: A Powerful Tool in Synthetic Biology

Wang, Hao; Wang, Lin; Zhong, Baihua; Dai, Zhuojun

doi:10.3389/fbioe.2022.810180

Cited by 24 publications

(15 citation statements)

References 71 publications

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“…When Cas9 are fused with effectors such as ABE, the size of the insert to be cloned is 5.3 Kb, thus making it impossible to clone in one single vector. Strategies aiming at splitting a large transgene into 2 smaller segments that can be cloned into individual AAV vector have recently emerged and include the overlapping, trans-splicing, hybrid or intein approaches (Patel et al, 2019;Wang et al, 2022). These strategies have been already applied on animal models for muscular dystrophies such as limb-girdle muscular dystrophy type 2B and Miyoshi myopathy or DMD (Lostal et al, 2010;Kodippili et al, 2018).…”

Section: Challenges Challenges Specific To Fshdmentioning

confidence: 99%

Gene Editing to Tackle Facioscapulohumeral Muscular Dystrophy

Mariot

Dumonceaux²

2022

Front. Genome Ed.

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Facioscapulohumeral dystrophy (FSHD) is a skeletal muscle disease caused by the aberrant expression of the DUX4 gene in the muscle tissue. To date, different therapeutic approaches have been proposed, targeting DUX4 at the DNA, RNA or protein levels. The recent development of the clustered regularly interspaced short-palindromic repeat (CRISPR) based technology opened new avenues of research, and FSHD is no exception. For the first time, a cure for genetic muscular diseases can be considered. Here, we describe CRISPR-based strategies that are currently being investigated for FSHD. The different approaches include the epigenome editing targeting the DUX4 gene and its promoter, gene editing targeting the polyadenylation of DUX4 using TALEN, CRISPR/cas9 or adenine base editing and the CRISPR-Cas9 genome editing for SMCHD1. We also discuss challenges facing the development of these gene editing based therapeutics.

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Section: Challenges Challenges Specific To Fshdmentioning

confidence: 99%

Gene Editing to Tackle Facioscapulohumeral Muscular Dystrophy

Mariot

Dumonceaux²

2022

Front. Genome Ed.

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“…The other method to expand the carrying capacity of AAVs is intein-mediated trans-splicing ( Tornabene et al, 2019 ). Split inteins are a pair of naturally occurring proteins that mediate protein trans-splicing in the same way that an intron in pre-mRNA splicing ( Wang et al, 2022 ). The split AAVs break a large transgene into two pieces and package each piece into an individual AAV.…”

Section: Introductionmentioning

confidence: 99%

Viral Vectors for the in Vivo Delivery of CRISPR Components: Advances and Challenges

Mengstie

2022

Front. Bioeng. Biotechnol.

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The Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) and its accompanying protein (Cas9) are now the most effective, efficient, and precise genome editing techniques. Two essential components of the CRISPR/Cas9 system are guide RNA (gRNA) and CRISPR-associated (Cas9) proteins. Choosing and implementing safe and effective delivery systems in the therapeutic application of CRISPR/Cas9 has proven to be a significant problem. For in vivo CRISPR/Cas9 delivery, viral vectors are the natural specialists. Due to their higher delivery effectiveness than other delivery methods, vectors such as adenoviral vectors (AdVs), adeno-associated viruses (AAVs), and lentivirus vectors (LVs) are now commonly employed as delivery methods. This review thoroughly examined recent achievements in using a variety of viral vectors as a means of CRISPR/Cas9 delivery, as well as the benefits and limitations of each viral vector. Future thoughts for overcoming the current restrictions and adapting the technology are also discussed.

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“…In this system, the C-terminal end of FucT2 and the N-terminal end of Oct-1 DNA-binding domain were fused to the N-terminal and C-terminal domains of an intein, respectively. Inteins are protein segments capable of mediating protein trans -splicing, which refers to the ligation of flanking extein into a new protein [ 19 ]. The efficiency of protein trans -splicing and FucT2 display on a plasmid were designed to be dependent on FucT2 solubility.…”

Section: Introductionmentioning

confidence: 99%