A CRISPR-Cas9-based transcription activation system was developed to enable efficient, genome-scale gene activation. The system, called SAM (Synergistic Activation Mediator), combines engineered dCas9 with multiple activation domains (e.g., VP64, p65, HSF1) and uses protein-interacting aptamers to enhance transcriptional activation. This system was tested on 12 genes and 6 long intergenic non-coding RNAs (lincRNAs), demonstrating robust activation with single-guide RNAs (sgRNAs). The system was also used to screen for genes that confer resistance to the BRAF inhibitor PLX-4720, identifying key resistance genes such as BCAR3, EGFR, and GPR35. The SAM system showed high specificity and efficiency, with activation levels up to 15-fold higher than previous methods. It was further validated using RNA sequencing and pharmacological assays, revealing that SAM-mediated activation is highly effective and specific. The system was also used to perform a genome-scale screening, identifying genes involved in BRAF inhibitor resistance. The results demonstrate the potential of SAM as a powerful tool for genome-scale gain-of-function screening in functional genomics research.A CRISPR-Cas9-based transcription activation system was developed to enable efficient, genome-scale gene activation. The system, called SAM (Synergistic Activation Mediator), combines engineered dCas9 with multiple activation domains (e.g., VP64, p65, HSF1) and uses protein-interacting aptamers to enhance transcriptional activation. This system was tested on 12 genes and 6 long intergenic non-coding RNAs (lincRNAs), demonstrating robust activation with single-guide RNAs (sgRNAs). The system was also used to screen for genes that confer resistance to the BRAF inhibitor PLX-4720, identifying key resistance genes such as BCAR3, EGFR, and GPR35. The SAM system showed high specificity and efficiency, with activation levels up to 15-fold higher than previous methods. It was further validated using RNA sequencing and pharmacological assays, revealing that SAM-mediated activation is highly effective and specific. The system was also used to perform a genome-scale screening, identifying genes involved in BRAF inhibitor resistance. The results demonstrate the potential of SAM as a powerful tool for genome-scale gain-of-function screening in functional genomics research.