Shang W., et al. (2018) Genome-wide CRISPR screen identifies FAM49B as a key regulator of actin dynamics and T cell activation. Proc. Natl. Acad. Sci. U S A.115(17):E4051-E4060 Summary: Researchers identified a previously uncharacterized gene named FAM49B, which acts as negative regulator in T cell activation, through genome-wide CRISPR screening. Titanium Taq polymerase was used in a two nested PCRs for genomic DNA extracted from screened cells. And the PCR products were submitted for sequencing on an Illumina HiSeq.
Stratigopoulos G., et al. (2018) DMSO increases efficiency of genome editing at two non-coding loci. PLoS One. 13(6):e0198637. Summary: Researchers found that DMSO may increases efficiency of CRISP/Cas9 genome editing at non-coding DNA. Substitution efficiency increased up to 10-fold by treatment of human embryonic stem cells (ESC) with non-toxic levels of DMSO (1%) before CRISPR/Cas9 delivery. Titanium Taq polymerase was used for these two SNPs region amplification before Sanger sequencing.
Tanney JB., et al. (2017) Aspergillus subgenus Polypaecilum from the built environment. Stud. Mycol. 88:237-267. Summary: In this study, researchers isolate approximately 9 000 strains mycobiota of indoor dust collected from 93 buildings in 12 countries worldwide. And they identified 5 novel species of subgenus Polypaecilum based on colony morphology and internal transcribed spacer rDNA region (ITS) barcodes. They used Titanium Taq polymerase for the amplification of ITS, and conducted phylogenetic analyses using sequences of ITS to confirm the five species of subgenus Polypaecilum.
Vandeweyer D., et al. (2017) Metagenetic analysis of the bacterial communities of edible insects from diverse production cycles at industrial rearing companies. Int. J. Food Microbiol. 261:11-18. Summary: In this study, authors compared the bacterial community composition of mealworms and crickets from several production cycles and rearing companies. They found that: for cricket rearing, production cycles of constant and good quality in terms of bacterial composition can be obtained by different rearing companies. For mealworms however, more variation in terms of microbial quality occurs between companies. Titanium Taq polymerase was used for Metagenetic analysis.
Spalek K., et al. (2016) A common NTRK2 variant is associated with emotional arousal and brain white-matter integrity in healthy young subjects. Transl. Psychiatry. 6:e758 Summary: The neurotrophic tyrosine kinase receptor type 2 gene (NTRK2) has been associated with many psychiatric diseases. Researchers investigated the relation between genetic variability of NTRK2 and emotional arousal in healthy young subjects. Titanium Taq polymerase was used for array-based SNP genotyping to identified a NTRK2 SNP associated with emotional arousal.
Multiplex PCR関連
Stellrecht, K. A.,et al. (2013) Comparison of three real-time PCR for the quantification of polyomavirus BK. J. Clin. Virol. 56:354–359. Summary: Latent polyomavirus BK, if reactivated during immunosuppression, can cause complications for kidney transplant patients. Researchers compared three different multiplex real-time PCR assays to analyze samples from 69 renal transplant patients for BK viral load. Titanium Taq polymerase was used with BK Virus Primers ASR from Luminex (MC-RTx assay) to amplify BK virus sequences from the VP2/VP3 region. The MC-RTx assay using Titanium Taq was found to have the greatest sensitivity and highest performance characteristics among the assays tested.
Buelow, D. R., et al. (2013) Comparison of two multiplexed PCR assays for the detection of HSV-1, HSV-2, and VZV with extracted and unextracted cutaneous and mucosal specimens. J. Clin. Virol. 58:84–88. Summary: Varicella zoster virus (VZV) and Herpes Simplex Virus 1 and 2 (HSV-1, HSV-2) can cause life-threatening infections in immunocompromised individuals. The authors examined the performance of analyte specific reagents (ASR) that were multiplexed for simultaneous detection of VZV, HSV-1, and HSV-2 from cutaneous or mucosal lesion clinical specimens. Titanium Taq was used for multiplex real-time PCR with Luminex (EraGen) ASR primer sets (MultiCode HSV primers, MultiCode VZV primers, and control primers) and was compared with another multiplexed commercial ASR assay (Focus Diagnostics). Successful multiplex analysis of dermal specimens was reported for both ASR assays.
Hilgert, N., et al. (2008) Mutation analysis of TMC1 identifies four new mutations and suggests an additional deafness gene at loci DFNA36 and DFNB7/11. Clin. Genetics74:223–232.
Summary: To understand the genetic heterogeneity of deafness, researchers analyzed 51 families with a history of autosomal dominant non-syndromic hearing loss. The samples were screened for copy number variation in the known disease-causing gene Transmembrane channel-like gene 1 (TMC1) using multiplex amplicon quantification (MAQ) with Titanium Taq polymerase. Twelve test amplicons located in the TMC1 locus and 8 reference amplicons were successfully amplified simultaneously.
Jiang, X., et al. (2007) Effect of renin-angiotensin-aldosterone system gene polymorphisms on blood pressure response to antihypertensive treatment. Chin. Med. J.120(9):782–786. Summary: Researchers identified polymorphisms in genes in the renin-angiotensin-aldosterone system (RAAS) among a population of Chinese Han patients with essential hypertension treated with an antihypertensive agent. Titanium Taq polymerase was used in a multiplex PCR genotyping assay covering seven amplicons. Data were correlated with blood pressure response to therapy to reveal various genotypes that correlated with positive therapeutic response.
Brouwers, N., et al. (2006) Genetic risk and transcriptional variability of amyloid precursor protein in Alzheimer’s disease. Brain129:2984–2991.
Summary: This genetic variation study analyzed the incidence of genomic locus duplication and presence of mutations in the promoter region of the amyloid precursor protein (APP) gene. Titanium Taq polymerase was used for multiplex amplicon quantification (MAQ) PCR assays to assess the copy number of APP and 11 neighboring genes. A total of 37 different amplicons were amplified in each multiplex PCR assay.
数個の細胞の微量サンプルからゲノムを増幅しCGH-アレイ解析（GenomePlex Single Cell Whole Genome Amplification Kit、Sigma-Aldrich）に使用した例
Identification of small gains and losses in single cells after whole genome amplification on tiling oligo arrays
Jochen B. Geigl, et al. Nucleic Acids Res. (2009) Aug;37:e105.
血清などの臨床サンプルからHepatitis C Virusを検出、同定する際に使用された例
Detection and Quantification of Hepatitis C Virus (HCV) by MultiCode-RTx Real-Time PCR Targeting the HCV 3' Untranslated Region
Elizabeth K. Mulligan, et al. J. Clin. Microbiol. (2009) Aug;47:2635-2638.
参考文献
Kellogg, D. E. et al. (1994) BioTechniques16(6):1137－1137.
