Merck Mouse Whole Genome SAM CRISPRa Pooled Lentiviral Library Kit Puro

Mouse Whole Genome SAM CRISPRa Pooled Lentiviral Library Kit Puro

포장

• 40 vials (8×25 μL aliquots for each of the 5 kit components)

적용 분야

• CRISPR

배송 상태

• Dry ice

저장 온도

• −70 °C

제품 설명

The transcriptional activator VP64 fused to nuclease-dead SpCas9 (dCas9) forms a ribonucleoprotein complex (RNP) with the CRISPR guide RNA (gRNA).
The stem- and tetra-loop sequences in the gRNA scaffold have been modified into minimal hairpin RNA aptamers that selectively bind dimerized MS2 bacteriophage coat proteins. The MS2 coat protein is fused to the p65 subunit of NF-κB (NFκB) and the activation domain of human heat-shock factor 1 (HSF1).
Each guide RNA contains two aptamers, each capable of binding two MS2 coactivator proteins, effectively recruiting four coactivators for every CRISPR activator complex.

The dCas9-VP64, MS2-p65-HSF1 helper constructs, and gRNA are delivered as three separate plasmids or lentiviral particles.
Unlike arrayed gRNA libraries where each clone is in a separate well, pooled gRNA libraries contain thousands of individual gRNAs in a single tube, enabling efficient genome-wide screening without robotics or specialized equipment.

The SAM CRISPR library is provided in 3 pools, each containing approximately 23,500 gRNAs (3 gRNAs per RefSeqID).
The whole genome library includes about 70,000 gRNAs, targeting 19,000 unique gene symbols (including alternative isoforms encompassing 23,500 RefSeqIDs).

컨트롤 구성

• 100 non-targeting control gRNAs are included within each pool for baseline comparison in statistical analysis of gRNA frequency changes via deep sequencing.
• Negative control gRNAs have minimal homology with the target genome and remain stable during control treatments.
• Positive controls must be determined empirically based on the phenotype of interest.

These controls provide flexibility for experimental design and downstream data analysis.

제품 스펙