基因表达和基因沉默
调节基因表达是对控制生物过程的复杂遗传网络进行研究的研究人员所采用的最强大技术之一。基因调节实验尤其被广泛用于确定疾病治疗方案的有效性，包括从癌症到发育和神经退行性疾病等。
目前有大量的技术可用于有效调节细胞和组织中的基因表达。RNAi（RNA干扰）是一种自然的生物学机制，其中siRNA（小干扰RNA）双链体可诱导有效的靶向基因表达抑制。SigmaAldrich® Advanced Genomics正成为为研究人员开发商品化RNAi产品的领导者。
最近，CRISPR抑制（CRISPRi）和CRISPR激活（CRISPRa）的发展为研究人员提供了一套强大的新工具，让几乎无限的基因调控选项成为可能。通过利用来自SigmaAldrich® Advanced Genomics无与伦比的CRISPR产品线，研究人员能够对几乎任何基因实现准确、高效的敲减、敲除、激活或过表达。
相关技术文章
- We offer advanced design and quality synthesis for predesigned siRNA and siRNA libraries using the Rosetta siRNA Design Algorithm to reduce off-target effects and increase RNAi performance. The algorithm utilizes Position-Specific Scoring Matrices (PSSM) and knowledge of the siRNA seed region to predict the most effective and specific siRNA sequences for a target gene of interest.
- Mature microRNAs (miRNAs) are a class of naturally occurring, small non-coding RNA molecules, about 21-25 nucleotides in length. MicroRNAs are partially complementary to one or more messenger RNA (mRNA) molecules, and their main function is to downregulate gene expression in a variety of manners, including translational repression, mRNA cleavage, and deadenylation.
- A series of vectors to enable a successful implementation of your RNAi experiments.
- Through the powerful science of RNA interference, successful gene silencing can be achieved with the use of synthetic, small interfering RNA (siRNA). By targeting these small, synthetic oligos, typically 19-23 bases in length against a specific gene of interest researchers are identifying gene function, elucidating pathways, and screening for potential new drug targets.
- MISSION® Application Data. We are committed to providing you with helpful application notes and data for the MISSION® TRC shRNA libraries.
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相关实验方案
- Complete protocol for Lentiviral transduction of suspension cells. This protocol describes the use of MISSION TRC shRNA Lentiviral Particles for long term silencing and phenotypic observation of suspension cells.
- FACS (Fluorescence-Activated Cell Sorting) provides a method for sorting a mixed population of cells into two or more groups, one cell at a time, based on the specific light scattering and fluorescence of each cell. This method provides fast, objective, and quantitative recording of fluorescent signals from individual cells.
- Method for purification, reverse transcription and quantitative PCR for MicroRNAs using Mysticq reagents
- Experimentally determine the appropriate antibiotic concentration for selecting stable cell lines. This titration protocol can determine the lowest concentration of puromycin needed to efficiently select transduced cells.
- Preparation of the Lentiviral Transduction Particles Using Packaging Plasmid Mix
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RNAi - 基因调控照亮遗传路径
功能基因组学筛选是分子靶点发现研究必不可少的环节，科学家可依靠该技术揭示生物系统、通路和疾病背后的潜在机制。对这些复杂的分子互作网络的有效研究可通过使用shRNA文库进行可靠、高效和长期的基因敲减来实现。
shRNA和siRNA介导的细胞中基因沉默的机制流程示意图。
Sigma-Aldrich® Advanced Genomics是RNAi协会TRC1.5和TRC2集合的唯一指定供应商。这类慢病毒shRNA文库基因组覆盖率全面，提供包括全基因组、单独RNAi克隆/载体和基因家族集的多种产品形式。
CRISPRi和CRISPRa - 强大的基因抑制和激活
CRISPRi（CRISPR干扰）和CRISPRa（CRISPR激活）可在不改变潜在的DNA序列的情况下分别提供高效的基因沉默和激活。当用于大规模 LOF（功能丧失）和 GOF（功能获得）筛选中时，研究人员能够识别出其他方法经常遗漏的独特但功能相关的基因通路。
带有协同激活介导因子复合物（SAM）的CRISPRa和使用KRAB域的CRISPRi的示意图。
Sigma-Aldrich® Advanced Genomics提供了一整套用于基因敲减和过表达实验的优化CRISPRi和CRISPRa文库目前可提供CRISPRi和SAM CRISPRa混合的慢病毒文库，也可根据您的特定需求提供定制产品。
担心CRISPR不适合您的实验项目？连线咨询Sigma-Aldrich®Advanced Genomics专家。
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