Single-cell Sequencing-based Pre-implantation Genetic Testing-M (PGT-M) of the Heterozygous Mutations of PKD1 Gene

Abstract

Objectives: Polycystic kidney disease (PKD) is a common autosomal monogenic genetic disease. PKD1 mutation accounts for about 85% of ADPKD patients. Pre-implantation genetic testing-M (PGT-M for monogenic) is an important approach to prevent the transmission of genetic diseases from parents to the offspring. Design: In this study, We have identified the family linkage and mutation site in embryos with NGS-based SNP phasing and Sanger Sequencing.Methods: Multiple Annealing and Looping Based Amplification Cycles (MALBAC) method was employed to amplify the whole genome of trophoblast cells. Copy Number Variant (CNV), and single nucleotide polymorphism (SNP) were used to assess the embryo state. Results: In the eight embryos, Embryo 02 and Embryo 04 were removed from further analysis because of the Multiple chromosomes abnormal (2 of 8, 25%). Embryo 05, Embryo 06, Embryo 07, and Embryo 08 were judged as 46,XN,-15q(q23→qter,~31M,×1,mos*), 45,XN,-16(×1), 47,XN,+2(×3),-7p(pter→p14.3,~35M,×1,mos*), and 46,XN, +16(×3,mos*),-20p(pter→p11.23,~23M,×1,mos*),+22(×3,mos*), respectively (4 of 8, 50%). Meanwhile, Embryo 01 and Embryo 03 were judged as 46, XN (2 of 8, 25%). The results of SNP phasing and Sanger Sequencing suggested that Embryo 01 and Embryo 05 had none of PKD1 gene mutation. Limitations: Up to now, PGT-M is complicated and expensive. Meanwhile, PGT-M obtains the final diagnosis through invasive manipulation of embryos, which may bring adverse effects on offspringConclusion: NGS-based single-cell sequencing combined with CNV, Sanger Sequencing, and SNP phasing is a reliable testing system for PGT-M application. This work presented here would provide a detailed understanding of the NGS-based single-cell sequencing application in ADPKD.

Considerations on gravitational waves in higher-order local and non-local gravity

“…, half of them are canonical but the remaining half are ghosts. Under the above BRS transformation (14), we find…”

“…the action ( 17) is invariant under the BRS transformation (14). The corresponding BRS current is given by…”

“…From now on, we shall indicate by a capital F a Lagrangian density containing also the contribution of a potential V (φ) and by F (φ), f (R), or f (R, R) a function of such fields without scalar field potential. Modifications can be considered also in the framework of Teleparallel Equivalent of General Relativity (TEGR) [13,14]. In such a case, dynamics is given by the torsion scalar T .…”

Role of the remnant symmetries in gravitational theories based on absolute parallelism: a 2D standpoint

“…[3] for an up to date review), but only in recent years the attention was paid to more formal, conceptual and fundamental aspects within the theory. After the first inkling in relation with the number of extra DoF [4], the bulk of the discussion on this subject started to develop recently [5], [6], not without some disparity in the results.…”

Thick string-like braneworlds in f(T) gravity

“…In cosmology, f (T ) models furnish geometric solutions to the early inflation [19] and to the late acceleration phase [20,21]. In spite of the extra degrees of freedom present in the vielbeins [22], the linear perturbation upon the flat spacetimes has only two propagating degrees of freedom [23][24][25]. The modifications of the f (T ) gravity was also explored on black holes [26][27][28],…”