6 results about "Single-stranded binding protein" patented technology

This invention discloses direct amplification qPCR reagents, kits, and applications for whole blood samples. The amplification reagents include: a composite hot-start DNA polymerase system, a buffer system, a surfactant, an anti-contamination system, and an anti-inhibition enhancer system. The composite hot-start DNA polymerase system is obtained by mixing antibody-modified hot-start Taq enzyme and antibody-modified Taq1C2 mutant Taq enzyme in a predetermined ratio. The buffer system includes Tris-HCl, MgCl2, (NH4)2SO4, and KCl. The surfactants include Tween-20 and NP-40. The anti-contamination system includes UDG enzyme and dNTPs, wherein the dNTPs are composed of dATP, dGTP, dCTP, dTTP, and dUTP. The anti-inhibition enhancer system includes D-trehalose, L-carnitine, DMSO, BSA, and SSB single-chain binding protein.

This invention belongs to the field of gene editing technology and discloses an inosine-enriched adenine base editor off-target detection method (Ino-seq) and its applications. The method of this invention utilizes endonuclease V to specifically cleave inosine-containing DNA, combined with the stabilization of a thermostable single-stranded DNA-binding protein and the high affinity enrichment of streptavidin-biotin, to achieve direct capture of ABE enzymatic products. This method can simultaneously detect sgRNA-dependent and independent off-target effects, with systematic comparisons showing F1 scores of 0.892–0.931. Targeted deep sequencing validation showed an overall accuracy of 95.3%. Ino-seq can also detect endogenous genomic inosine, observing a variation range of >1,000-fold in 10 cell lines. This invention provides a crucial safety assessment tool for the clinical translation of ABE technology, supporting comprehensive off-target analysis in preclinical and clinical studies.

Provided are a fusion protein that improves gene editing efficiency and an application thereof. The fusion protein comprises a single-stranded DNA binding protein functional domain, nucleoside deaminase and nuclease. According to CBEs, when carrying our base conversion from C-G to T-A, nucleoside deaminase such as cytosine deaminase carries out deamination by using single-stranded DNA as a substrate, and by re-fusing the single-stranded DNA binding protein functional domain on the fusion protein of the nucleoside deaminase and nuclease, the chance of single-stranded DNA being exposed to the nucleoside deaminase is greatly increased, thereby significantly improving base editing efficiency. The present disclosure provides a breakthrough improvement of single-base gene editing technology and can greatly promote the application thereof in aspects such as gene editing, gene therapy, cell therapy, animal model making, and crop genetic breeding.

This invention belongs to the field of bioengineering and discloses a *Yersinia lipolytica* chassis strain with homologous recombination preference, its construction method, and its applications. It involves inserting endogenous genes Rad52, Rad50, Mus81, and Mms4, as well as the single-stranded DNA annealing protein (PapRecT) encoded by *Pseudomonas aeruginosa* bacteriophage and its corresponding host *P. aeruginosa*-derived single-stranded DNA binding protein (PaSSB), into the genome of the *Yersinia lipolytica* strain, followed by knocking out its endogenous Rad9, resulting in a *Yersinia lipolytica* chassis strain with homologous recombination preference. This invention's *Yersinia lipolytica* chassis strain achieves effective improvement in homologous recombination efficiency through multi-dimensional regulation of the homologous recombination process, constructing an excellent *Yersinia lipolytica* platform, which is beneficial for further genome editing and has significant implications for using *Yersinia lipolytica* as a chassis cell in basic or applied research.

The present invention provides genetically modified microorganisms with reduced transmissibility, wherein the microorganism has been genetically modified to express a single stranded binding protein (SSB) regulated by an arabinose responsive promoter, wherein the microorganism has a reduced growth capacity in the absence of arabinose.

This invention relates to the field of molecular biology detection technology, specifically, to the application of a fluorescent fusion protein in the detection of DNA damage repair. The application includes: incubating the fluorescent fusion protein with a test sample and then performing fluorescence imaging to obtain the damage repair status of the DNA to be tested in the test sample; wherein the test sample contains the DNA to be tested; and the fluorescent fusion protein is formed by the fusion of a single-stranded DNA-binding protein and a fluorescent protein. This application not only enables in-situ detection of any DNA damage that exposes single-stranded DNA, but also provides accurate and quantifiable results. Furthermore, this invention is not limited to any species and can provide efficient visualization and quantitative analysis for DNA damage detection in various organisms.