4 results about "Cloning" patented technology

Gating clocks has been a widely adopted technique for reducing dynamic power. The clock gating strategy employed has a huge bearing on the clock tree synthesis quality along with the impact to leakage and dynamic power. This invention is a technique for clock gate optimization to aid the clock tree synthesis. The technique enables cloning and redistribution of the fanout among the existing equivalent clock gates. The technique is placement aware and hence reduces overall clock wire length and area. The technique involves employing the k-means clustering algorithm to geographically partition the design's registers. This invention improves the clock tree synthesis quality on a complex design.

The invention belongs to the technical field of biological preparation and in particular relates to an RXRalpha (Homo Sapiens Retinoid X Receptor alpha) gene knockout cell system with stable and low expression of an RXRalpha protein and a preparation method of the RXRalpha gene knockout cell system. The cell system is an RXRalpha gene knockout cell system; the knockout base number is an integral multiple of a number except 3; RXRalpha protein expression in a cell is remarkably reduced, the genotype is stable and can be passed to next generations, the stability of the RXRalpha low-expression cell system is greatly improved, and stable generation passage and continuous culture of the cell system can be facilitated. The invention provides the preparation method of the cell system. The preparation method comprises the following steps: according to RXRalpha gene sequence information, designing a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) knockout gRNA (Guide Ribonucleic Acid) sequence, establishing a gRNA expression vector, and carrying out in-vitro cell level detection on gRNA shearing activity; by using a nuclear transferring method, carrying out cotransfectionon an immortalized cell of human neuroblastoma, that is, an SK-N-SH cell, by using cas9 and the gRNA expression vector, carrying out G418 medicine screening so as to obtain stable cell cloning, carrying out a PCR (Polymerase Chain Reaction) and gene sequencing to identify that the cell is cell cloning of which the RXRalpha gene is knocked out for an integral multiple of a number except 3, therebyobtaining the RXRalpha gene knockout cell system with stable and low expression of the RXRalpha protein.