8 results about "CRISPR/Cpf1" patented technology

CRISPR / Cpf1-related compositions and methods for treatment of cancer.

To provide a composition and method for modifying primary T cells.SOLUTION: The present invention is a method of gene editing comprising administering stem-loop CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) RNA (st-crRNA) and Cpf1 enzyme to a cell.SELECTED DRAWING: None

The present invention discloses a method for improving the acetate tolerance and lipid accumulation of oil-producing microorganisms by using acetyl-CoA synthetase (ACS), belonging to the technical fields of genetic engineering and microbial engineering. In the present invention, the endogenous constitutive MaACS is overexpressed in Mortierella alpina, improving the acetate tolerance, lipid production and the proportion of polyunsaturated fatty acids (PUFAs) represented by arachidonic acid (ARA) of the recombinant strain, and a medium with ammonium acetate as the nitrogen source is used to further improve its lipid accumulation. The present invention also further optimizes the acetyl-CoA metabolic pathway by using the CRISPR / Cpf1 multi-gene operating system, and co-expresses citrate lyase (ACL1) while overexpressing the MaACS to further strengthen the supply of acetyl-CoA, or expresses glucose-6-phosphate dehydrogenase (G6PD2) to balance the supply of reducing power NADPH to avoid the spillage of acetyl-CoA, or knockdowns the SNF1γ subunit of the sucrose non-fermenting protein kinase complex that negatively regulates lipid accumulation to improve the activity of acetyl-CoA carboxylase 1 (ACC1).

This invention pertains to recombinant AsCpf1 and LbCpf1 nucleic acids and polypeptides for use in CRISPR / Cpf1 endonuclease systems and mammalian cell lines encoding recombinant AsCpf1 or LbCpf1 polypeptides. The invention includes recombinant ribonucleoprotein complexes and CRSPR / Cpf1 endonuclease systems having a suitable AsCpf1 crRNA is selected from a length-truncated AsCpf1 crRNA, a chemically-modified AsCpf1 crRNA, or an AsCpf1 crRNA comprising both length truncations and chemical modifications. Methods of performing gene editing using these systems and reagents are also provided.

The invention relates to bacillus subtilis for producing D-psicose and application of the bacillus subtilis, and belongs to the technical field of microorganisms. The invention discloses a recombinant bacillus subtilis obtained by performing gene editing by using a CRISPR / Cpf1 system, a D-psicose 6-phosphoric acid 3-epimerase coding gene A6PE, a glucokinase coding gene glck and a glucose transporter coding gene glf are integrally expressed in a starting bacillus subtilis genome, and a 6-phosphofructokinase coding gene pfkA is knocked out, so that the recombinant bacillus subtilis is obtained. A metabolic pathway for producing D-psicose by taking glucose as a substrate is constructed in bacillus subtilis, and the production efficiency is improved in a form of enhancing glucose uptake and conversion capacity, so that the yield of D-psicose of the constructed recombinant bacillus subtilis at a 250 mL shake flask fermentation level reaches 23 g / L. The bacillus subtilis for producing D-psicose provided by the invention has the advantages of directional regulation and control of metabolic flux, high substrate utilization rate, low production cost and the like, and is suitable for industrial production.

The present invention discloses a method and strain for optimizing the production of N-acetylglucosamine using multiple pathways. The present invention is based on Bacillus subtilis 168 strain, and uses a genome editing system based on CRISPR / Cpf1 to construct a synthetic strain of N-acetylglucosamine from scratch. The competitive pathway is optimized by a high-throughput combination of a period-type CRISPRi system based on a stable phase promoter and dCpf1, thereby obtaining a more efficient and stable N-acetylglucosamine synthetic strain. The gene knockout and promoter replacement of the recombinant Bacillus subtilis constructed by the present invention are both completed using a genome scarless editing system based on CRISPR / Cpf1, and no plasmid is used in the final strain. Therefore, glucose can be sustainably, stably and efficiently synthesized into acetylglucosamine, and its yield on a shake flask can reach 33.7g / L, laying the foundation for its industrialization.

This invention pertains to recombinant AsCpf1 and LbCpf1 nucleic acids and polypeptides for use in CRISPR / Cpf1 endonuclease systems and mammalian cell lines encoding recombinant AsCpf1 or LbCpf1 polypeptides. The invention includes recombinant ribonucleoprotein complexes and CRSPR / Cpf1 endonuclease systems having a suitable AsCpf1 crRNA is selected from a length-truncated AsCpf1 crRNA, a chemically-modified AsCpf1 crRNA, or an AsCpf1 crRNA comprising both length truncations and chemical modifications. Methods of performing gene editing using these systems and reagents are also provided.

CRISPR / Cpf1-related compositions and methods for treatment of cancer.