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Point Mutation Cell Line

Point Mutation Cell Line

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The Bingo™ Prime Editing platform, a novel system for CRISPR point mutation cell line construction, is exclusively developed by EDITGENE. With this novel platform, we provide accurate and efficient CRISPR Cas9 point mutation service. Bingo™ platform is based on the most efficient and accurate point mutation technology - Prime Editing (PE). EDITGENE has over a decade of gene editing experience, and we have summarized and optimized the targeting strategy design principle from hundreds of gene-editing projects. The positive rate is far higher than that of the traditional HDR-based point mutation system.


Five optimized aspects of cripsr point mutation cell line


Features


There are over 75000 diseases and genetic related mutations in human, and single point mutations are accounting for the largest proportion. However, there are still no good treatment methods for most of them so far. CRISPR point mutation cell line is an excellent tool for the research of both genetic diseases and cancers diagnosis and treatment.

At present, the most common point mutation method is based on CRISPR/Cas gene editing technology, which can generate DSBs (Double-strand breaks) at specific locus, so that exogenous donor oligo can be integrated into the genome through homologous recombination to achieve the purpose of knockin. However, the off-target effect caused by CRISPR/Cas system has been an issue in gene editing, and the introduction of double-strand breaks has also caused problems such as byproducts and gene transpose. Therefore, the application of homologous recombination based gene therapy is limited.

The gene editing technology without double-strand breaks can only achieve 4 types of base editing (C→T, G→A, A→G, T→C), and there are still 8 kinds of editing that cannot be achieved (C→A, C→G, G→C, G→T, A→C, A→T, T→A, T→G). Leveraging Cas enzyme, reverse transcriptase and prime editing guide RNA (pegRNA), the prime editing system breaks through the above limitations and achieves precise gene editing for any kinds of base mutation.

The BingoTM point mutation cell line service provided by EDITGENE is based on the prime editing system, which can accurately and efficiently introduce the target mutation to cells. The BingoTM Prime Editing platform helps to promote the development of disease research and treatment.

●   Cell type: Cancer cell lines, Stem cells and unlimited-passage cells that can be transfected.

  EDITGENE has handling experience of over 300 types of cell lines. View the cell list

●   Price: Precise price depends on cell types and project difficulty, please contact our specialist for a 

quote, or email us for any query (info@editxor.com)

●   Turnaround: As fast as 10 weeks. In-stock point mutation cell line deliver in one week.

 

Services:

crispr cas9 point mutation services


Why EDITGENE?


Workflow


crispr cas9 point mutation workflow

1. Design a pegRNA with a transcription template (carrying the desired point mutation) and the necessary Prime Binding Site (PBS).

crispr cas9 point mutation workflow

2.Under the guidance of pegRNA, Cas9 cleaves single-stranded DNA, and the PBS at the 3' terminus of pegRNA can recognize and pair with the complementary sequence before the cutting site.

crispr cas9 point mutation workflow

3. Reverse transcriptase (M-MLV RT) uses the designed template sequence behind the PBS sequence on the pegRNA as the template for reverse transcription, and directly polymerizes the target sequence onto the cut DNA.

crispr cas9 point mutation workflow

4. The edited DNA double-strand extends a DNA Flap, activating the cell's 3' flap - 5' flap acquisition mechanism to remove the original DNA fragment.

crispr cas9 point mutation workflow

5.Unmatched DNA double strands are repaired through a mismatch repair (MMR) mechanism to repair non-edited strands, and the entire editing process is completed.


Case Study


Case 1: CTNNB1 c.98C>T mutation cell line (HCT 116)

Result: The sequencing data shows that the mutation knockin was success.